Sinh Hoc Phan Tu

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B Y T
SINH HC PHN T
(DNG CHO O TO DC S I HC)
M s: .20.X.06
NH XUT BN GIO DC
Ch o bin son:
V KHOA HC V O TO - B Y T
Ch bin:
GS.TS. NGUYN VN THANH
Nhng ngi bin son:

GS.TS. NGUYN VN THANH
TS. TRN THU HOA
TS. TRN CT NG
ThS. H TH YN LINH
Tham gia t chc bn tho:

ThS. PH VN THM
TS. NGUYN MNH PHA
file://C:\Windows\Temp\nlddyqkdsp\sinh_hoc_phan_tu.htm 30/09/2009
Bn quyn thuc B Y t (V Khoa hc v o to)
770 - 2007/CXB/3 - 1676/GD M s: 7K721M7 - DAI
Li gii thiu
Thc hin mt s iu ca Lut Gio dc, B Gio dc & o to v B Y t ban hnh chng
trnh khung o to Dc s i hc. B Y t t chc bin son ti liu dy - hc cc mn c s v
chuyn mn theo chng trnh trn nhm tng bc xy dng b sch t chun chuyn mn trong
cng tc o to nhn lc y t.
Sch SINH HC PHN T c bin son da trn chng trnh gio dc ca Trng i hc Y
Dc Thnh ph H Ch Minh trn c s chng trnh khung c ph duyt. Sch c cc tc gi
GS.TS. Nguyn Vn Thanh, TS. Trn Thu Hoa, TS. Trn Ct ng, ThS. H Th Yn Linh bin son theo
phng chm: Kin thc c bn, h thng; ni dung chnh xc, khoa hc; cp nht cc tin b khoa
hc, k thut hin i v thc tin Vit Nam.
Sch SINH HC PHN T c Hi ng chuyn mn thm nh sch v ti liu dy - hc

chuyn ngnh Dc s i hc ca B Y t thm nh vo nm 2007. B Y t quyt nh ban hnh l ti
liu dy - hc t chun chuyn mn ca ngnh trong giai on hin nay. Trong thi gian t 3 n 5
nm, sch phi c chnh l, b sung v cp nht.
B Y t xin chn thnh cm n cc tc gi v Hi ng chuyn mn thm nh gip hon thnh

cun sch; Cm n PGS.TS. Nguyn Vn Ty, PGS.TS. inh Hu Dung c v phn bin, cun
sch hon thnh kp thi phc v cho cng tc o to nhn lc y t.
Ln u xut bn, chng ti mong nhn c kin ng gp ca ng nghip, cc bn sinh vin

v cc c gi ln xut bn sau c hon thin hn.
V KHOA HC V O TO - B Y T
LI NI U
Nm 1909, Johansen W. xut bn chuyn kho "Cc yu t ca hc thuyt ng n v bin d v di
truyn" (Element de exakten Erblichkeitslehre), trong ln u tin xut hin t gen. Nm 1953,
Watson v Crick khm ph ra m hnh xon kp ADN thc y nhanh chng s pht trin ca di
truyn hc mc phn t. Nm 1965, J. Watson xut bn sch "Sinh hc phn t ca gen" dy 494
trang v n nm 1976, trong ln ti bn ln th ba dy ln 739 trang. Tip sau , hng lot ti liu
v Sinh hc phn t ra i.
Cho n ngy 26 - 06 - 2000, ti Washington D.C, cng ty t nhn Celera Genomics (Anh) v D n
B gen Ngi (Human Genome Project) ca Vin nghin cu Quc gia v Sc khe ca Hoa K
(National Institute of Health) phc tho bn b gen ngi. Theo , b gen ngi c 3,12 t
nucleotid v 97% tng nucleotid c xc nh trnh t, trong c 85% s trnh t t ng v
tr. Khong 3% ADN c cha gen, 97% cn li l ADN "khng chc nng". Trong tng s 3% ADN
ny c khong 30 - 50000 gen.
Ngy 14 - 4 - 2003, T chc Quc t nh trnh t B gen Ngi (International Human Genome
Sequencing Consortium) tuyn b hon thnh nhng cng on cui cng ca bn gen ngi.
K bn gen, cc phng php tm gen c tnh cht tr liu s c thc hin quy m ln v
Dc l b gen (Pharmacogenomics) s khm ph su hn b gen ngi ng dng trong ngnh
Dc.
Sch "Sinh hc phn t" nhm gip cho sinh vin Dc khoa hiu c cu trc c bn v chc
nng ca gen. Sch gm cc bi: Nhp mn; Sao chp ADN; Cc loi ARN; S phin m v m di
truyn; Sinh tng hp protein; iu ho hot ng gen; B gen t bo nhn tht; t bin gen - Cc
phng php phn tch ADN.
Sch xut bn ln u nn kh trnh khi thiu st. Cc tc gi rt mong nhn c s gp ca

c gi.
CC TC GI
CC T VIT TT
A Adenine
ADN Acid desoxyribonucleic
AIDS Acquired immune deficiency syndrome
AP Apurinic or apyrimidinic
ARN Acid ribonucleic
bp Base pair
C Cytosine
cADN Complementary DNA
cds Coding sequence
CpG C phosphat G
CTF CCAAT binding Trascription Factor
DMD Duchenne muscular dystrophy
DNase Deoxyribonuclease
dNTP Desoxyribonucleozid triphosphat
dsADN Double stranded DNA
eEF Eukaryote elongation factor
EF Elongation factor
eIF Eukaryote initiation factor
eRF Eukaryote release factor
EGF Epidermal growth factor
FRET Fluorescence resonance energy transfer
G Guanine
GMO Genetic modified organism
Hfr High frequency recombination
Hft High frequency transduction
HIV Human immunodeficiency virus
hnARN Heterogenous nuclear RNA
Icr Insensitive to catabolite repression
IF Initiation factor
IGS Internal guide sequence
Kb Kilobase
LINE Long interspersed repetitive element
MALDI-TOF Matrix assisted lazer desorption ionization time of
fligt
mARN Messenger RNA
NAAAF N acetoxy 2 acetylaminofluorene
NAD Nicotinamide adenine dinucleotide
NER Nucleotide excision repair

NF Neurofibromatosis
NF1 Neurofibromatosis type 1
NMN Nicotinamide mononucleotide
NMP Nucleotide monophosphat
Ori Origin of replication
PCR Polymerase chain reaction
PDGF Platelet derived growth factor
PFGE Pulse field gel electrophoresis
Pi Inefficient promoter
PKU Phenylketonuria
PR Photoreactivation
Rad Radiation absorbed dose
rARN Ribosome RNA
RE Restriction enzyme, restriction endonuclease
Rem Roentgen equivalent man
RF Release factor
RFI Replicative form I
RFLP Restriction fragments length polymorphism
RNase Ribonuclease
RNP Ribonucleoprotein
RRF Ribosome release factor
scRNA Small cytoplasmic RNA
SINE Short interspersed repetitive element
snRNA Small nuclear RNA
SPR Surface plasmon resonace
SRP Signal recognition particle
SSB Single stranded binding
ssADN Single stranded DNA

T Thymine
tRNA Transfer RNA
THE Transposable human element
THR Thyroid Hormon Receptor
Tm Melting temperature
U Uracile
UTR Untranslated region
BNG I CHIU THUT NG VIT - ANH

Ting Vit Ting Anh
ADN b sung Complementary DNA
ADN si i Double - stranded DNA
ADN si n Single - stranded DNA
ADN vi v tinh Microsatellite DNA
Alkapton niu Alkaptonuria
ARN nhn khng ng nht, hnARN Heterogenous nuclear RNA
ARN nhn nh, snARN Small nuclear RNA
ARN polymerase ph thuc ADN DNA - dependent RNA polymerase
ARN ribosom Ribosomal RNA
ARN t bo cht nh, scARN Small cytoplasmic RNA
ARN thng tin Messenger RNA
ARN vn chuyn Transfer RNA
Base ng ng Base analogue
Bnh hng ban lupus Systemic lupus erythematosus
Bnh lon dng c Duchenne Duchenne muscular dystrophy
Bin np Transformation
B ba kt thc Stop codon
B gen Genome
B suy gim Attenuator
Bong bng sao chp Replication bubble
Chc ba sao chp Replicating fork
Cht cm ng Inducer
Cht d nhim sc Heterochromatin
Cht d nhim sc c cu Constitutive heterochromatin
Cht hot ho Activator
Cht nguyn nhim sc Euchromatin
Cht nhim sc Chromatin
Cht siu c ch Superrepressor
Cht c ch Inhibitor, repressor
Cht c ch gc Aporepressor
Chip ADN DNA microarray
Chui xon kp Double helix
Chuyn v nghch Retrotransposition
C ch bn bo tn Semiconservative mechanism
C ch bo tn Conservative mechanism
C ch ln vng Rolling circle mechanism
Cng ngh di truyn Genetic engineering
Cng ngh sinh hc Biotechnology
Cng ngh nano sinh hc Bionanotechnology
Du n ADN DNA fingerprint
D xc tc Heterocatalysis
D n b gen ngi Human genome project
a cistron Polycistron
a kim sot Multiple control
u dnh Cohesive end
u t Blunt end
im gc sao chp Origin of replication
in di gel Gel electrophoresis
in di trng xung
Pulse field gel electrophoresis
iu ho m Negative control
iu ho dng Positive control
nh kiu gen Genotyping
on chn Insertion sequence
on d Probe
on Okazaki Okazaki fragment
i m Anticodon
n cistron Monocistron
n v phin m Operon
n v sao chp Replicon
t bin cm ng Induced mutation
t bin cng ngha Sense mutation
t bin lng Silent mutation
t bin lch ngha Missense mutation
t bin t nhin Spontaneous mutation
t bin v ngha Nonsense mutation
Enzym ct gii hn Restriction enzyme
Enzym phin m ngc Reverse transcriptase
Gy t bin im nh hng Site - directed mutagensis
Gen c th cm ng Inducible gene
Gen c th c ch Repressible gene
Gen iu ho Regulatory gene
Gen gi Pseudogene
Gen nhy Transposon
Gen tin ung th Proto - oncogene
Gen ung th Oncogene
Hch nhn Nucleolus
Ht F F - prime
B gen hc Genomics
H protein Proteome
H protein hc Proteonomics
Heterogenous nuclear ARN hnRNA
Ho min dch Immunochemistry
Hc thuyt trung tm Central dogma
Hp Pribnow Pribnow box
Hp TATA TATA box
Hp t Zygote
Hp t khng hon ton Merozygote
Kh np Competence
Khun mu Template
Kim sot m Negative control
Kim sot cm ng m Negative inducible control
Kim sot cm ng dng Positive inducible control
Kim sot dng Positive control
Kim sot sau dch m Post - translation control
Kim sot c ch m Negative repressible control
Kiu gen Genotype
Kiu hnh Phenotype
K thut di truyn Genetic engineering
Lai ti ch In situ hybridization
Lai vt ADN Southern blot
Lai vt ARN Northern blot
Lai vt protein Western blot
Lng ghp Catena
Lc lp Chloroplast
Lc tip hp Matting force
M di truyn Genetic code
Mng t bo cht Cell membrance
Mi Primer
Mi trng ti thiu Minimal media
Ngoi nhim sc th Extrachromosome
Ngn tay km Zinc finger
Nhn con Nucleolus
Nhim sc th Chromosome
Nhim sc th a si Polytene chromosome
Nicotinamide mononucleotide NMN
Nicotinamide - adenine dinucleotide NAD
Nt Loop
Nt kp tc Hairpin loop
Operon c th cm ng Inducible operon
Phage n ho Temperate phage
Phn ca gen cu trc khng mang m Intron
Phn ca gen cu trc mang m Exon
Phn tha u Terminal redundancy
Phn ng chui polymerase Polymerase chain reaction
Phenyl ceton niu Phenylketonuria
Phc hp khi ng ng Closed promoter complex
Phc hp khi ng m Open promoter complex
Phc hp nhn din tn hiu xut Signal recognition particle
Promoter v tinh Spacer promoter
Protein cng mch Single - stranded binding protein
Protein iu ho Regulatory protein
Protein gn chp Cap - binding protein
Protein hiu ng Effector protein
Protein c ch Repressor protein
Quang phc hi Photoreactivation
Sn phm khuch i Amplicon
Sau dch m Post - translation
Sinh hc phn t Molecular biology
Sinh vt chuyn gen Genetic modified organism
Si sm Leading strand
Si khun sm Leading strand template
Si khun mun Lagging strand template
Si mun Lagging strand
S bin hnh d lp th Allosteric transformation
S ct ni Splicing
S a hnh di cc on ct gii hn Restriction fragments length polymorphism
S d bit codon Codon bias
S h bin Tautomerization
S phin m Transcription
S sao chp Replication
S t ct ni Self - splicing
S t iu ho Autoregulation
S c ch ngc Feedback inhibition
Sa cha bng cch ct nucleotid Nucleotide excision repair
Tc nhn gy t bin Mutagen
Ti np Transduction
T bo cho Donor
T bo nhn Recipient
T bo nhn nguyn thu Prokaryote
T bo nhn tht Eukaryote
Thnh t bo Cell wall
Th ct ni Spliceosome
Th a hnh Polymorphism
Th khuyt dng Auxotroph
Th nguyn dng Prototroph
Th nhn Nucleoid
Th sao chp Replicative form
Th sinh dng Soma
Th th Receptor
Thc khun th Bacteriophage
Tin thc khun Prophage
Tin virus Provirus
Tip hp Conjugation
Tin sinh hc Bioformatics
T chc hch nhn Nucleolus organizer
Trnh t chung Consensus sequence
Trnh t iu ho Operator
Trnh t hng dn ni ti Internal guide sequence
Trnh t kt thc Terminator
Trnh t m ho Coding sequence
Trnh t tng cng Enhancer sequence
Ung th gic mc di truyn Retinoblastoma
V tr AP AP site
V tr iu ho Regulatory site
V tr khi u Initiator site
V virus Capsid
Vng khi ng Promoter
Vng khng dch m Untranslated region
Vng khng m ho Noncoding region
Vng m ho Coding region
Yu t di truyn ng nghch Retroposon, retrotransposon
Yu t di truyn vn ng Transposable genetic element
Yu t gii tnh Sex - factor
Yu t ko di Elongation factor
Yu t kt thc Release factor
Yu t khi u Initiation factor
Yu t phng thch ribosom Ribosome release factor
Yu t tng cng Enhancer
Yu t tng trng thng b Epidermal growth factor
Yu t tng trng t tiu cu Platelet - derived growth factor
BNG I CHIU THUT NG ANH - VIT
Ting Anh Ting Vit
Activator Cht hot ho
Alkaptonuria Alkapton niu
Allosteric transformation S bin hnh d lp th
Amplicon Sn phm khuch i
Anticodon i m
AP site V tr AP
Aporepressor Cht c ch gc
Attenuator B suy gim
Autoregulation S t iu ho
Auxotroph Th khuyt dng
Bacteriophage Thc khun th
Base analogue Base ng ng
Bioformatics Tin sinh hc
Bionanotechnology Cng ngh nano sinh hc
Biotechnology Cng ngh sinh hc
Blunt end u t
Cap Chp
Cap - binding protein Protein gn chp
Capsid V virus
Catena Lng ghp
Cell membrance Mng t bo cht
Cell wall Thnh t bo
Central dogma Hc thuyt trung tm
Chloroplast Lc lp
Chromatin Cht nhim sc
Chromosome Nhim sc th
Closed promoter complex Phc hp khi ng ng
Coding region Vng m ho
Coding sequence Trnh t m ho
Codon bias S d bit codon
Cohesive Dnh
Cohesive end u dnh
Competence Kh np
Complementary DNA ADN b sung
Conjugation Tip hp
Consensus sequence Trnh t chung
Conservative mechanism C ch bo tn
Constitutive heterochromatin Cht d nhim sc c cu
Corepressor Cht ng c ch
De novo Mi
DNA fingerprint Du n ADN
DNA microarray Chip ADN
DNA - dependent RNA polymerase ARN polymerase ph thuc ADN
Donor T bo cho
Double helix Chui xon kp
Double - stranded DNA ADN si i
Duchenne muscular dystrophy Bnh lon dng c Duchenne
Effector protein Protein hiu ng
Elongation factor Yu t ko di
Enhancer Yu t tng cng
Enhancer sequence Trnh t tng cng
Epidermal growth factor Yu t tng trng thng b
Euchromatin Cht nguyn nhim sc
Eukaryote T bo nhn tht
Exon Phn ca gen cu trc mang m
Extrachromosome Ngoi nhim sc th
Feedback inhibition S c ch ngc
F - prime Ht F
Gap Khong trng
Gel electrophoresis in di gel
Genetic code M di truyn
Genetic engineering Cng ngh di truyn, k thut di truyn
Genetic modified organism Sinh vt chuyn gen
Genome B gen
Genomics B gen hc
Genotype Kiu gen
Genotyping nh kiu gen
Hairpin loop Nt kp tc
Heterocatalysis D xc tc
Heterochromatin Cht d nhim sc
Heterochromatin Cht d nhim sc
Heterogenous nuclear RNA ARN nhn khng ng nht, hnARN
Human genome project D n b gen ngi
Immunochemistry Ho min dch
In silico Trn my tnh
In situ Ti ch
In situ hybridization Lai ti ch
In vitro Trong ng nghim
In vivo Trn sinh vt
Induced mutation t bin cm ng
Inducer Cht cm ng
Inducible gene Gen c th cm ng
Inducible operon Operon c th cm ng
Inhibitor Cht c ch
Initiation factor Yu t khi u
Initiator site V tr khi u
Insertion sequence on chn
Internal guide sequence Trnh t hng dn ni ti
Intron Phn ca gen cu trc khng mang m
Lagging strand Si mun
Lagging strand template Si khun mun
Leading strand Si sm
Leading strand template
Si khun sm
Loop Nt
Matting force Lc tip hp
Merozygote Hp t khng hon ton
Messenger RNA ARN thng tin
Microsatellite DNA ADN vi v tinh
Minimal media Mi trng ti thiu
Missense mutation t bin lch ngha
Molecular biology Sinh hc phn t
Monocistron n cistron
Multiple control a kim sot
Mutagen Tc nhn gy t bin
Negative control iu ho m, kim sot m
Negative inducible control Kim sot cm ng m
Negative repressible control Kim sot c ch m
Noncoding region Vng khng m ho
Nonsense mutation t bin v ngha
Northern blot Lai vt ARN
Nucleoid Th nhn
Nucleolus Hch nhn
Nucleolus organizer T chc hch nhn
Nucleotide excision repair Sa cha bng cch ct nucleotid
Okazaki fragment on Okazaki
Oncogene Gen ung th
Open promoter complex Phc hp khi ng m
Operator Trnh t iu ho
Operon n v phin m
Phage Thc khun
Phenotype Kiu hnh
Phenylketonuria Phenyl ceton niu
Photoreactivation Quang phc hi
Platelet - derived growth factor
Yu t tng trng t tiu cu
Polycistron a cistron
Polymerase chain reaction Phn ng chui polymerase
Polymorphism Th a hnh
Polytene chromosome Nhim sc th a si
Positive control iu ho dng, kim sot dng
Positive inducible control Kim sot cm ng dng
Post - translation Sau dch m
Post - translation control Kim sot sau dch m
Pribnow box Hp Pribnow
Primer Mi
Probe on d
Prokaryote T bo nhn nguyn thu
Promoter Vng khi ng
Prophage Tim thc khun
Proteome H protein
Proteonomics H protein hc
Proto - oncogene Gen tin ung th
Prototroph Th nguyn dng
Provirus Tin virus
Pseudogene Gen gi
Pulse field gel electrophoresis in di trng xung
Receptor Th th
Recipient T bo nhn
Regulatory gene Gen iu ho
Regulatory protein Protein iu ho
Regulatory site V tr iu ho
Release factor Yu t kt thc
Replicating fork Chc ba sao chp
Replication S sao chp
Replication bubble Bong bng sao chp
Replicative form Th sao chp
Replicon n v sao chp
Repressible gene Gen c th c ch
Repressor Cht c ch
Repressor protein Protein c ch
Restriction enzyme Enzym ct gii hn
Restriction fragments length polymorphism S a hnh di cc on ct gii hn
Retinoblastoma Ung th vng mc
Retroposon Yu t di truyn ng nghch
Retrotransposition Chuyn v nghch
Retrotransposon Yu t di truyn ng nghch
Reverse transcriptase Enzym phin m ngc
Ribosomal RNA ARN ribosom
Ribosome release factor Yu t phng thch ribosom
Rolling circle mechanism C ch ln vng
Self - splicing S t ct ni
Semiconservative mechanism C ch bn bo tn
Sex - factor Yu t gii tnh
Signal recognition particle Phc hp nhn din tn hiu xut
Silent mutation t bin cng ngha, t bin ln
Single - stranded binding protein Protein gn si n
Single - stranded DNA ADN si n
Site - directed mutagensis Gy t bin im nh hng
Small cytoplasmic RNA ARN t bo cht nh, scARN
Small nuclear RNA ARN nhn nh, snARN
Soma Th sinh dng
Southern blot Lai vt ADN
Spacer promoter Promoter v tinh
Spliceosome Th ct ni
Splicing S ct ni
Spontaneous mutation t bin t nhin
Stop codon B ba kt thc
Superrepressor Cht siu c ch
Systemic lupus erythematosus Bnh hng ban lupus
TATA box Hp TATA
Tautomerization S h bin
Temperate phage Phage n ho
Template Khun mu
Terminator Trnh t kt thc
Transcription S phin m
Transduction Ti np
Transfer RNA ARN vn chuyn
Transformation Bin np
Transposable genetic element Yu t di truyn vn ng
Transposon Gen nhy
Untranslated region Vng khng dch m
Western blot Lai vt protein
Zinc finger Ngn tay km
Zygote Hp t
Bi 1
NHP MN SINH HC PHN T
MC TIU
- Trnh by c lch s v phng php nghin cu sinh hc phn t.
- Nu c mc tiu v i tng mn hc.
- Trnh by c cc thnh tu hin i do sinh hc phn t em li.
1.1. LC S
C th phn chia s pht trin ca sinh hc phn t thnh ba giai on chnh.
1.1.1. Giai on hnh thnh cc tin
Friedrich Miescher khm ph acid nucleic vo nm 1869, khi ng ch l mt bc s 22 tui. Khi

thu phn m bnh nhn bng pepsin v acid hydrochloric ri chit vi ether, ng ta thu c nhn t
bo. Khi x l nhn t bo vi kim, ng thu c mt cht ta m ng gi l nuclein v do tnh cht
acid ca n nn c gi l acid nucleic. V sau Miescher s dng tinh trng ca c hi chit acid
nucleic cho nghin cu ca mnh.
Gregor Mendel khm ph quy lut di truyn nm 1865, khi nghin cu s truyn tnh trng trn cy
u H Lan.
Vo u th k XX, Walter Sutton v Theodor Boveri thit lp mi quan h gia Di truyn hc

v Sinh hc khi quan st s phn ly ca nhim sc th t bo. Nhng v nhim sc th cha c protein
ln acid nucleic nn ngi ta bt u nghin cu xc nh u l vt liu di truyn tht s.
Frederick Griffith l mt nhn vin y t London vi nhim v chnh l nghin cu cc cn dch.

Nm 1928 nghin cu s nhim ph cu trong i dch cm sau Th chin I, trong ghi nhn s
chuyn i kiu hnh t R sang S ca ph cu c th xy ra ngay c khi dng t bo cht.
Nm 1944, Oswald Avery, Colin MacLeod, v Maclyn McCarty cng b bng chng thc
nghim u tin rng chnh ADN l vt liu di truyn. Avery v cc cng s c bi bo ca Griffith v
h c lp li th nghim vi nh tm kim cht chu trch nhim cho s chuyn i nhng khng
thnh cng. Sau nh mt phng php tch loi protein ra khi dung dch ca M. G. Sevag h
thnh cng trong vic chng minh ADN chnh l tc nhn truyn tnh trng.
Nm 1951, Erwin Chargaff, chng minh trong ADN t l nucleotid Adenin lun bng Thymin;
Cytosin bng Guanin hay ni cch khc t l base purin bng t l base pyrimidin; trong khi t l A + T
G + C v thay i theo loi.
Nhng n nm 1952, vai tr di truyn ca ADN mi c xc nhn. Khi Hershey v Chase s

dng k thut nh du phng x chng minh ADN ch khng phi protein l cht mang thng tin di
truyn.
1.1.2. Giai on Sinh hc phn t ra i
Nm 1953, m hnh cu trc ADN ca Watson - Crick ra i, c xem l khm ph ln nht trong
Sinh hc ca th k. M hnh xon kp ADN do James D. Watson v Francis H. C. Crick xy dng l s
kt hp ca cng trnh v t l nucleotid do Edwin Chargaff xng v cng trnh nghin cu si
ADN bng tn x tia X ca Rosalind Elsie Franklin v Maurice Wilkins.
Nm 1961, M. Nirenberg v J. Matthei tm ra b m di truyn nhn to u tin. n gia nhng

nm 1961, ton b 64 codon (b ba m ho) c xc nh.
c bit nm 1961, F. Jacob v J. Monod tm ra c ch di truyn iu ho tng hp protein.
Nm 1962, Warner Arber pht hin ra enzym ct gii hn trong t bo vi khun.
Nm 1967, enzym ADN ligase c chit xut. Ngi ta xem enzym ny l keo dn phn t, c th
ni hai si ADN li vi nhau.
Nm 1970, Hamilton Smith chit c enzym ct gii hn.
1.1.3. Sinh hc phn t hin i
Trong thp nin 70 ca th k XX, k thut di truyn ra i to nn cuc cch mng mi trong di
truyn v sinh hc phn t. Vic xc nh trnh t nucleotid (ADN sequencing) ca gen nhanh chng
dn n k thut mi: gy t bin im nh hng cho php to cc bin i tu trn gen.
Nm 1973, A.C. Chang v Herbert Boyer, to c nhng plasmid ti t hp u tin.

Phng php ny c m rng vo nm 1973 v ngi ta ni nhiu on ADN vo plasmid SC101
tch ra t E. coli. Cc phn t ti t hp
ny c th t sao chp khi a vo t bo E. coli khc. Nh vy Sinh hc phn t (SHPT) thc y s
ra i ca mt ngnh cng ngh mi l cng ngh di truyn.
Nm 1985, R.K. Saiki v K. B. Mullis cng b vic s dng PCR khuch i cc on gen
- globin in vitro. T y thao tc ADN (ADN manipulation), xc nh trnh t gen pht trin rng ri
v ng dng trong nhiu lnh vc khc nhau nh chn on, bin i di truyn, xc nh ph h,
C th ni SHPT trong giai on hin i c p dng trong ba lnh vc ch yu:
- Nghin cu c bn v cu trc v chc nng ca tng gen.
- Sn xut protein hu ch bng phng php mi.
- To ra cc sinh vt bin i gen (GMO).
Vic hon thnh bn gen ngi gip gii quyt nhng vn ln nh iu tr ung th, pht
trin phi, bit ho t bo.
Vo u nm 1990, s kt hp gia Sinh hc phn t v Tin hc a n mt phng php mi l

in silico (nghin cu sinh hc trn my in ton).
Bng 1.1. Cc ct mc quan trng trong s hnh thnh v pht trin ca SHPT
Thi gian S kin
1830 Pht hin protein.
1833 Pht hin v phn lp enzym u tin.
1865 Di truyn hc ra i: Gregor Mendel khm ph quy lut di truyn cc tnh

trng.
1869 Friedrich Miescher khm ph acid nucleic
1879 Fleming pht hin cht nhim sc.
1900 Drosophila (rui gim) c s dng trong cc nghin cu gen.
1906 Thut ng "Di truyn hc" ln u tin c s dng.
1911 Rous pht hin virus gy ung th u tin.
1915 Pht hin thc khun (phage).
1919 Ln u tin t "Cng ngh sinh hc" (biotechnology) c dng.
1938 Thut ng "Sinh hc phn t" (molecular biology) c a ra bi Warren

Weaver.
1941 Thut ng "Cng ngh di truyn" (genetic engineering) c dng ln u,

bi nh vi sinh vt hc ngi H Lan A. Jost trong bi ging ti hc vin k
thut Lwow, Ba Lan.
1944 Oswald Avery v cng s chng minh ADN mang thng tin di truyn.
1946 Pht hin rng vt liu di truyn t cc virus khc nhau c th phi hp to
ra virus mi, mt th d v ti t hp di truyn.
1947 McClintock khm ph yu t di truyn vn ng "gen nhy" bp ng.
1949 Pauling chng minh bnh hng cu li lim l mt "bnh phn t" do t
bin trong protein hemoglobin.
1953 James Watson v Francis Crick cng b cu trc xon kp ADN, m ra thi
i mi ca sinh hc.
1955 Phn lp c enzym chu trch nhim tng hp acid nucleic.
1956 Kornberg khm ph ADN polymerase I, dn n vic lm sng t c ch sao

chp ADN.
1958 Chng minh bnh hng cu li lim ch do t bin 1 acid amin.
ADN c tch ra trong ng nghim ln u tin.
1959 Cc bc trong qu trnh sinh tng hp protein c phc ha.
1960 S dng tnh cht b sung cp base to phn t ARN - ADN lai.
Tm thy ARN thng tin.
1961 - 1966 Khm ph m di truyn gm cc b ba (codon).
1967 My nh trnh t protein t ng u tin hon chnh.
1969 Tng hp enzym in vitro ln u tin.
1970 Pht hin enzym gii hn, m ng cho vic to dng gen.
1971 Tng hp c gen hon chnh u tin.
1973 Stanley Cohen v Herbert Boyer ct v ni ADN thnh cng (dng enzym gii
hn v ligase) v to ra ADN mi vi khun.
1976 Cc cng c ti t hp ADN ln u tin c p dng cho ngi ri lon di

truyn.
Lai phn t c p dng chun on tin sinh bnh alpha thalassemia.
Gen ca nm men c biu hin E. coli.
Trnh t ca mt gen c xc nh.
1977 Ln u tin biu hin gen ngi vi khun.
Pht trin k thut nh trnh t nhanh cc ADN di nh in di.
1978 Xc nh cc cu trc chi tit ca virus.
Chng minh kh nng a t bin im vo v tr c hiu trn ADN.
Thp nin 70 Pht hin cc polymerase.
Hon chnh k thut nh trnh t acid nucleic.
nh hng gen.
Ct ni ARN.
1980 Trao bng sng ch k thut to dng cho Cohen v Boyer.
Pht trin my tng hp gen u tin.
Gii Nobel Ho hc cho k thut ti t hp phn t: Berg, Gilbert, Sanger.
1982 Applied Biosystems, Inc., gii thiu my nh trnh t protein pha kh u tin,
gim thiu lng protein cn nh trnh t.
1983 Tng hp nhim sc th nhn to u tin.
Cc nh v di truyn u tin cho cc bnh di truyn c pht hin.
1984 Pht trin k thut du n ADN.
Ton b b gen ca HIV c to dng v nh trnh t.
1985 Tm thy cc nh v di truyn ca bnh thn v x nang.
PCR c pht minh. V tr thnh cng c chnh trong nghin cu v pht

trin sn phm khp th gii.
Kt qu du n di truyn u tin c s dng tranh tng ti to.
1988 Quc hi M thng qua Human Genome Project.
1989 Bt u Plant Genome Project.
Thp nin 80 Cc nghin cu ADN c dng xc nh lch s tin ho.
Pht hin ribozyme v retinoblastoma.
1990 Human Genome Project c khi ng.
1994 Gen ung th v u tin c pht hin.
1995 Trnh t b gen hon chnh ca mt sinh vt khng phi virus c xc nh:
Hemophilus influenzae.
1996 Pht hin gen lin quan n bnh Parkinson.
1998 B gen ca ng vt u tin, giun C. elegans, c nh trnh t.
Bn s b ca b gen ngi c cng b cho thy c hn 30000 gen.
Thp nin 90 Bn n u tin c tuyn da vo bng chng du n di truyn Anh
2000 Bn gen hon chnh ca thc vt u tin: Arabidopsis thaliana.
Cng b bn s b ca b gen ngi.
2001 Hon chnh bn gen cy lng thc u tin: cy la.
Hon chnh bn gen ca vi khun nng nghip: Sinorhizobium meliloti, vi

khun c nh m v Agrobacterium tumefaciens, thuc dit su ry thc
vt.
2002 Bn proteom s b u tin ca nm men.
Thnh lp t hp quc t nh trnh t cc k sinh trng gy st rt v mui

st rt.
Cc nh khoa hc lm sng t cc yu t kim sot s bit ho t bo gc,

xc nh trn 200 gen lin quan.
2003 Cng b phin bn th ca b gen ngi hon chnh.
2004 FDA ln u tin cng nhn h thng ADN microarray, Ampli Chip
Cytochrome P450 Genotyping Test, gip chn on bnh.
Gii m B gen G.
20/10/2004 Cng b bn m t khoa hc hon chnh ca B gen Ngi cho thy c

khong 20000 - 25000 gen m ho cho protein
1.2. I TNG V PHNG PHP NGHIN CU
1.2.1. nh ngha
Sinh hc phn t l mt b phn ca Sinh hc, khoa hc v s sng, vi i tng nghin cu l s

sng cp phn t, tp trung vo cc kha cnh v cu trc, s sao chp v biu hin ca gen; s
tng tc v chc nng sinh l ca cc sn phm ca gen. Cc khoa hc sinh hc khc nh Di truyn
hc hay Ho sinh cng nghin cu sinh hc mc phn t, tuy nhin thin v chc nng sinh hc
ca gen hay sn phm ca gen hn l chnh cc phn t ny (hnh 1.1).
Hnh 1.1. Quan h gia sinh hc phn t vi cc khoa hc sinh hc khc
1.2.2. Hc thuyt trung tm
Hc thuyt trung tm (Central Dogma) ca Sinh hc phn t c pht biu ln u tin bi Francis
Crick nm 1958 v s lun chuyn thng tin ca sinh vt: "Thng tin khi chuyn sang protein th
khng th ly ra li c". Ngha l thng tin khng th chuyn ngc t protein n acid nucleic.
Thng tin y l trnh t chnh xc cc nucleotid ca acid nucleic quy nh trnh t acid amin ca
protein.
Hnh 1.2. S lun chuyn thng tin ca sinh vt
Theo thng tin c th c chuyn gia cc phn t ADN thng qua s sao chp hoc gia phn
t ADN v ARN thng qua s phin m hoc t acid nucleic n protein nh s dch m. Ngoi ra
thng tin cn c th di chuyn gia ARN n ARN trong qu trnh sinh sn ca cc ARN virus hay qu
trnh x l ct ni ca ARN hoc ngc li t ARN n ADN trong qu trnh phin m ngc ca
retrovirus. Cc qu trnh ny nghin cu chi tit trong Sinh hc phn t.
1.2.3. Cc phng php nghin cu
T cui nhng nm 50 n u nhng nm 60 ca th k XX, cc nh Sinh hc phn t c c cc

k thut phn lp, xc nh c tnh v thao tc trn cc phn t sinh hc nh ADN, ARN v protein.
Cc k thut ny bao gm:
1.2.3.1. To dng biu hin
To dng biu hin c s dng nghin cu chc nng protein. on ADN cn quan tm c
to dng vo mt plasmid v a vo t bo biu hin. Bng cch thay i c cu biu hin trn
plasmid hoc tinh ch protein ngi ta c th hiu c cch thc iu ho hot ng ca gen, quan h
ca n vi hot ng ca cc gen hay protein khc cng nh cu trc v chc nng ca n.
1.2.3.2. PCR
PCR l mt k thut cho php khuch i mt on gen c hiu thu c nhiu bn sao phc v
nghin cu. Bn cnh , PCR cng cho php d dng a cc thay i, t bin vo ADN nghin
cu.
1.2.3.3. in di gel
y l mt cng c phn tch chnh trong Sinh hc phn t. N da vo vic phn tch cc phn t
sinh hc nh ADN, ARN, protein trong in trng, trong cc phn t ny s di chuyn di tc
ng ca in trng xuyn qua mt h gel, do chng c th c tch ra theo kch thc hay in
tch.
1.2.3.4. Lai vt Southern v Northern
K thut lai vt Southern c s dng pht hin s hin din ca mt trnh t ADN c hiu
trong mu bng cch chuyn cc phn t ADN c phn tch bng in di gel sang mt mng rn
v cho lai pht hin nh mt on d c hiu c nh du. K thut lai vt Northern vi nguyn l
tng t nhng c p dng cho i tng ARN, c s dng nghin cu s biu hin ca cc
ARN.
1.2.3.5. Lai vt Western v ho min dch
Protein cng c th c phn tch trn gel v lai vt trn mng rn (lai vt Western) v pht hin
bng k thut ho min dch. Nh c th nghin cu s hin din ca protein trong t bo v lin h
vi s biu hin ca gen tng ng.
1.3. NHNG NG GP LN CA SINH HC PHN T HIN NAY
1.3.1. Genomics: gii m b gen v ngnh h gen hc
Genomics (b gen hc) l khoa hc nghin cu mt cch h thng cc trnh t ADN y

(genome) ca sinh vt. Genomics thit lp thng tin v s sng bng cc tip cn mt cch h thng v
c th tin hnh quy m cng nghip. Genomics khc vi genetics (di truyn hc). Genetics nhm vo
cc gen n l, ti mt thi im, nh l chp nh nhanh. Genomics c gng nhm vo tt c cc gen
nh mt h ng hc, qua thi gian, xc nh chng tng tc nhau v nh hng nh th no n
cc con ng sinh hc, cc mng li v sinh l trong ng cnh tng qut hn. Nh vy, genomics
chnh l mt nhnh khoa hc gii thng tin mt m trong ADN v khm ph cc thng tin nh s lng
gen, t chc v ni dung ca b gen. Thng tin ny c p dng cc k to ln trong Y t, Nng nghip,
Sinh hc tin ho v cc lnh vc khoa hc khc.
C nhiu loi genomics tu theo kha cnh v lnh vc m n nghin cu p dng. Trong mi loi
genomics cng li gm nhiu lnh vc, v d nh plant genomics gm cng ngh sinh hc thc vt, sinh
hc phn t v cc k thut lin quan n thc vt (nng nghip, lm nghip, lm vn).
Kh nng gii trnh t y b gen sinh vt y mnh vic xc nh chc nng cc gen mc
ngha rng ca b gen. V cc gen c chc nng lin quan c iu ho cng nhau, cc k thut
nh gi s biu hin ton b gen s gip nhn nh c ch khi u v cm cc trnh t gen mi c
chc nng lin quan. Xp cc trnh t gen thnh cc nhm chc nng theo s biu hin ca gen s cho
hng dn c bn cc thc nghim b sung, nhm xc nh c im chc nng chnh xc ca sn phm
cui cng ca gen. Trong hai thp k va qua, cc k thut nh gi s biu hin gen c nhng
tin b, t cc phng php trn cc gen n l c hiu (v d lai vt Northern, slot v dot; phin m
ngc v PCR bn nh lng v nh lng; phng php bo v nuclease) n cc k thut tp trung
vo nhn nh tt c cc gen c biu hin khc nhau gia hoc trong cc mu th nghim.
Cc phng php nhn nh cc khc bit ca biu hin gen gm c lai loi tr (subtractive
hybridization), trnh din khc bit (differential display), phn tch hng lot biu hin gen (SAGE,
serial analysis of gene expression) v lai vi bn tri (microarray hybridization).
Trc y, theo phng php th cng, mi tun, mi ngi ch thc hin c mt vi phn ng
gii trnh t vi nng sut 300 bp/phn ng. Ngy nay vi h thng my mao mch c th xc nh t
ng ng thi 96 phn ng vi di trn 1000 bp/phn ng. Nh n gii trnh t b gen ngi
di 3,2 t nucleotid hon thnh vo thng 4/2003.
99% b gen ngi c c trnh t vi ni dung chnh xc 99,99%. Thnh tu v gii m b

gen ngi v nhiu sinh vt khc nh rui gim, giun trn, chut v cc vi sinh vt khc nh E. coli,
Saccharomyces cerevisae, C. briggsae, Drosophila pseudobsura v gn y ca cy la nc, m ra
mt ngnh khoa hc mi l "Genom hc - Genomics" chuyn nghin cu v cu trc v chc nng ca
ton b b gen sinh vt.
1.3.2. Proteomics: phn tch bin ng protein v ngnh h protein hc
Proteomics l nghin cu quy m ln v protein nhm thit lp cc du hiu nhn dng, s lng
cu trc v chc nng sinh ho v t bo ca tt c protein trong c th, c quan hoc tiu c quan
cng nh s thay i cc c tnh ny theo khng gian, thi gian v trng thi sinh l. T "proteomics"
c t ra tng ng vi genomics. Proteome (b protein) ca mt sinh vt l tp hp cc protein
bi n trong thi gian sng ca n. Proteome ca mt t bo tp hp cc protein trong n. T
"proteome" xut pht t "proteins" v "genome", v genome (b gen) m ho cc protein.
Proteomics thng c xem l bc tip theo trong nghin cu cc h thng sinh hc, sau
genomics. N phc tp hn genomics nhiu, ch yu l v b gen ca sinh vt n nh hn, proteome
khc nhau gia cc t bo v thay i lin tc qua cc tng tc sinh ho ca n vi b gen v mi
trng. Mt sinh vt c s biu hin protein khc nhau hon ton cc phn khc nhau trong c th ca
n. Mt kh khn chnh na l tnh phc tp ca cc protein lin quan n cc acid nucleic.
Kin thc v proteomics s gip hiu sinh vt tt hn nhiu so vi genomics. Cc phng php nh
phosphoproteomics v glycoproteomics c dng nghin cu cc bin i sau dch m (post -
translational modifications). Cc nghin cu proteomics bao gm nghin cu tng tc ca cc protein
v nhn din protein.
1.3.2.1. Tng tc ca cc protein
Hu ht cc protein hot ng hp tc vi cc protein khc v mt trong nhng ch ca proteomics

l nhn din tng tc ca cc protein. y thng l u mi quan trng v chc nng ca cc protein
mi tm ra. c mt vi phng php d cc tng tc protein - protein. Phng php kinh in l
phn tch lai i trn nm men (yeast two - hybrid analysis). Cc phng php mi gm c vi bn tri
protein (protein microarrays), sc k i lc min dch (immunoaffinity chromatography), tip theo l
khi ph (mass spectrometry), v kt hp cc phng php thc nghim nh trnh din phage v cc
phng php my tnh.
1.3.2.2. Nhn din protein
Nghin cu proteomics hin nay trc ht l protein phi c phn gii, i khi quy m ln. C
th tch protein bng in di hai chiu trn gel (two - dimensional gel electrophoresis). K thut ny u
tin tch protein theo im ng in v sau theo khi lng phn t. Cc im protein trn gel c
th nhn c bng cch dng cc cht nhum ho hc khc nhau hoc cc du hiu hunh quang.
Thng th cc protein c th nh lng da vo cng nhum mu ca chng. Cc protein c
nhn din khi c tch ring v nh lng. Cc m ring r c ct ra khi gel v ct thnh cc
peptid bng cc enzym thu phn gii protein. Sau cc peptid ny c nhn din bng khi ph, c
bit l khi ph MALDI - TOF. Trong quy trnh ny, peptid c t trn matrix lm peptid to cc tinh
th. Sau peptid c ion ho vi tia laser v s tng in th matrix c dng bn cc ion v
pha u d (detector) m thi gian ion n u d tu thuc vo khi lng ca n. Ion c khi lng
cng cao cng mt nhiu thi gian bay. Trong khi ph MALDI - TOF, cc ion cng c th b lch
hng vi gng phn x tnh in cng lm tp trung chm ion. Nh vy, c th xc nh vi chnh
xc cao s n u d th hai cc khi ca cc ion v cc khi ny c th pht hin chnh xc cc thnh
phn ca cc peptid, do nhn din chng.
Cc hn hp protein c th c phn tch khng cn tch trc. Quy trnh ny bt u bng phn
gii cc protein trong phc hn hp. Cc peptid to thnh thng c bm vo ct sc k lng hiu
nng cao (HPLC) lm tch cc peptid da vo tnh khng a nc. Cc peptid tch ra t ct c th
c nhn din bng khi ph song song (MS/MS). Giai on u tin ca khi ph song song l phn
lp cc ion peptid ring r, giai on th hai s ph v cc peptid thnh cc on v s dng khun mu
ca on (fragmentation pattern) xc nh trnh t acid amin ca chng. C th dng nh du vi
cc nhn ng v so snh nh lng nng cc protein trong hai hoc nhiu hn hai mu
protein.
1.3.2.3. Cc k thut c s dng trong proteomics
- in di mt chiu v hai chiu trn gel (One - and two - dimensional gel electrophoresis).
- Tinh th tia X v cng hng t ht nhn (X - ray crystallography and nuclear magnetic
resonance).
- a khi ph (Tandem mass spectrometry) kt hp vi sc k o pha (reverse phase

chromatography) hoc in di hai chiu (2 - D electrophoresis).
- Khi ph (khng song song), thng l MALDI - TOF.
- Sc k i lc (Affinity chromatography), k thut lai i trn nm men (yeast two hybrid

techniques), chuyn nng lng cng hng hunh quang (FRET), v cng hng plasmon b mt
(SPR) c dng nhn nh tng tc protein - protein v lin kt protein - ADN.
- X - ray Tomography.
- Phn mm.
Vi cc k thut sc k, in di, trc y ngi ta ch nghin cu c tng loi protein ring r.

Ngy nay, khi phi hp sc k a chiu v khi ph ngi ta c th phn tch cng lc 5000 loi protein
v kt qu cho php chn on sm nhng bnh him ngho nh ung th mu. Proteomics nhm tin ti
gii m chc nng sinh hc ca h gen, song trc mt ch mi nghin cu c nhng bin i hot
ng ca cc nhm gen trong iu kin bnh l, cung cp thng tin cho vic chn on sm, phng tr
v iu tr nhiu loi bnh khc nhau.
1.3.3. Genomics, proteomics v s pht trin thuc
Pht minh v pht trin thuc l mt qu trnh di, sn phm ra th trng cn khong 15 nm t
khi c tng cho thuc c ch tim nng. Cc giai on c bn qu trnh c minh ha trong hnh
1.3. Di truyn hc c th tc ng n tng pha ca qu trnh ny. Tc ng ca di truyn hc v b gen
hc ln x hi s l d on di truyn v an ton v hiu qu ca thuc sm nht trong thng
trng.
Hnh 1.3. Qu trnh pht minh v pht trin thuc

v cc c hi tham gia ca genomics, proteomics
1.3.3.1. Pharmacogenomics v chin lc pht trin thuc
Pharmacogenomics (dc l b gen) nghin cu s nh hng ca di truyn ln p ng ca c th

vi thuc. T ny xut pht t pharmacology v genomics v nh vy n l s giao nhau ca dc v di
truyn.
Genomics to ra s thay i c bn trong cch s dng thuc v qu trnh pht trin thuc. S thay
i quy m ny c l s xy ra trong 5 n 10 nm ti. Nguyn nhn ca s thay i l cuc cch mng
khoa hc v k thut trong lnh vc B gen Ngi (Human genome, bn a hnh n nucleotid
(single - nucleotide polymorphism, SNP), nh loi gen (genotyping) v tin sinh hc (bioinformatics).
SNP l nhng du hiu o lng v th nghim tm sot hiu nng cao. Cc tin b trong nh
gi biu hin gen cng l diu k. Trong vng nm nm, s lng cc gen, nhy ca cc th nghim,
lp li ca cc kt qu cng gia tng ng k kh nng phn tch cc s liu. o s biu hin gen
c dng trong di truyn ung th v dc hc b gen ung th (cancer pharmacogenomics). S biu
hin gen gip cch phn loi ung th v liu php thch hp tt hn.
Cc li ch m pharmacogenomics em li gm c:
- Cho cc thuc cng hiu hn. Cc thuc mi da trn cc phn t protein, enzym, v RNA lin
quan n cc gen v bnh c ch to. C th pht minh thuc v cho cc du hiu thuc (drug maker)
a ra liu php c ch chuyn bit. S chnh xc ny khng ch ti a ho hiu qu tr liu m cn
lm gim s ph hu cc t bo khe mnh gn .
- Cho cc thuc tt hn, an ton hn ngay t u. Thay v phng php chun th - v - sai (m
mm) cc bnh nhn ph hp vi cc thuc ng t ban u, cc bc s c th phn tch lc di
truyn ca bnh nhn v k toa thuc tt nht c. y khng ch l cng vic tm thi tm thuc
ng, m cn cho thi gian bnh phc nhanh v tng an ton v cc phn ng ph c th xy ra s
c hn ch.
- Xc nh liu thuc thch hp bng phng php chnh xc hn. Cc phng php hin nay
nh liu c s theo khi lng v tui s c thay th bng nh liu c s theo di truyn ca con
ngi c th x l thuc v thi gian chuyn ho n nh th no. iu ny s ti a ho gi tr ca
liu php v cn gim kh nng qu liu xy ra.
- Tin xa trong sng lc bnh. Kin thc m di truyn ngi s cho php con ngi to li sng
thch hp v cc thay i mi trng sm trnh hoc gim mc trm trng ca bnh di truyn.
Tng t nh vy, tin b trong hiu bit tnh nhy cm ca bnh, c bit s cho php a ra bin php
theo di v iu tr cn thn nhng giai on thch hp nht ti a ho liu php cho chng.
- Ci tin trong pht minh, pht trin thuc. Cc qu trnh thch hp lm d dng th nhm trn
nhm dn s c di truyn chuyn bit cho mc thnh cng ln hn. Gi thnh v nguy c ca cc
th nghim lm sng gim do ch nhm vo nhng ngi c th p ng vi thuc.
- Gim chi ph trong chm sc sc khe. Gim s lng cc phn ng ph ca thuc, s lng th
thuc tht bi, thi gian thuc c ph duyt, thi gian bnh nhn cn dng thuc, s lng thuc
m bnh nhn phi dng iu tr hiu qu, nh hng ca bnh ln c th (do pht hin sm) v tng
khong cch ch c th c ca thuc s y mnh gim mng li gi thnh trong chm sc sc

khe.
1.3.3.2. Chin lc chemogenomics pht minh thuc
Cc thnh tu trong genomic v proteomic nhn din cc ch mi cho s can thip ca thuc l
cc c hi cha tng c cho pht minh cc tc nhn mi c cc kiu liu php tc ng mi.
S tip cn ca "chemogenomics" nh gi ch dng thng tin c bn cung cp bi trnh t ch

lm protein v ri sau pht minh "hp cht cng c" phn t nh tng tc vi ch. Hp cht
cng c c th c nh gi trong m hnh bnh th trc tip gi thuyt tr liu. S tip cn ny c
th c thc hin nh mt qu trnh song song v c bit thch hp pht minh thuc trong nhng
h ln. Mt v d ca chin lc chemogenomics phn lp cc ch phn t c phc tho trong
hnh 1.4.
Hnh 1.4. Mt v d thc tin v kinh t ca chin lc chemogenomics
Trnh t gen ca cc ch c xc nh bng cc tip cn genomics c to dng v biu hin

thnh cc protein ch (a, b) v cc protein ny thch hp cho vic sng lc vi mt th vin mu d
gm cc hp cht ging thuc (c). Cc hp cht ny c sng lc tm cc phn t c hot tnh bng
cch s dng mt th nghim lin kt c tnh ph qut v nh lng (d). Cc phn t c hot tnh ban
u hay cc d liu nh lng v hot tnh - cu trc pht sinh t th nghim lin kt c phn tch (e)
v s dng thit lp chin lc chn lc cho s tng hp mi cc hp cht vi cc c tnh c ci
tin. Cc hp cht ny c chn t mt c s d liu cc ng ng c th tng hp c ca th vin
mu d ban u (f) c tng hp bng cc phng php tng hp song song (g) v c th nghim
thit lp profile hot tnh - cu trc ca ch ang nghin cu v tinh chnh cc tiu ch sng lc cc
vng tip theo. Trong mi chu k, u th c gn cho cc ng vin tng hp bng cch s dng
cc qu trnh ti u ho a bin c thit k nhm m bo cc hp cht khng ch c ti u ho v
i lc gn vi ch m cn c cc c tnh ging thuc cho php chng c s dng trc tip nh l
cc hp cht mu trong cc m hnh sinh hc hay t bo thch hp (h).
1.3.4. Sn xut v s dng chip ADN
Chip ADN c hnh dng ging nh mt con chip vi nhng chm ADN thay cho transitor. L mt
mnh mng lin kt cao c kch thc 20 40 mm c in trn cc on ADN. V d chip b gen
ngi c in 20000 - 50000 gen nhng im chm vung cc nh.
Khi lai chip ADN ny vi sn phm phin m ca b gen c th, cc chm ADN i mu tng ng
vi mc hot ng ca nhng gen tng ng trong c th trng thi v thi im nghin cu. T
kt lun c tnh trng bnh l ca bnh nhn.
Chip ADN ang dn dn tr thnh cng c chn on trong cng nghip ln men vi sinh, trong y
hc d phng, trong kim dch v an ton thc phm v trong kim sot mi trng.
1.3.5. Chuyn gen vo cy trng
Ngi ta thc hin chuyn vo cy trng cc loi gen tng cng kh nng khng su bnh nh gen
khng su nhm oxy/VIP, gen khng virus nhm CP/Nbi, gen khng thuc dit c nhm bar. Nhng
cng trnh chuyn gen l vo thuc l cho thy thc vt c biu hin gen l ging nh E.coli. Vo nhng
nm 1996, cc nh cng nghip (Monsanto) a ra th trng nhng sn phm cy chuyn gen u
tin nh bp (ng), u nnh v bng vi. Nm 1996 trn th gii mi c 0,5 ha cy trng chuyn gen
c a vo sn xut. n nay (2003) din tch trng cy chuyn gen trn quy m ton cu ln ti
67 triu ha. Ngy nay, ngi ta ang tm cch a gen sn xut vaccin, gen sn xut dc cht (cc
pepetid nh), cc cht dinh dng (vitamin A trong ht la), cy trng t lng thc thc phm tr
thnh cy sn xut dc liu c gi tr kinh t cao.
Trong s 50 loi cy trng mang gen l ang c th nghim th cc cy bng vi khng su, cy
u tng, cy ng khng su, khng thuc dit c chim tng s trn 90% din tch.
1.3.6. Tin sinh hc
Cng ngh tin sinh hc bao gm cc phng php khai thc nhanh ngn hng d liu, phn tch
trnh t v cu trc ADN v protein. Tin sinh hc ang ci tin phng php x l phn tch s liu, ci
thin kh nng d on vng hot ng, vng ngng ngh ca b gen, ci tin kh nng phng on
phn ng t bo i vi tc nhn ngoi sinh, thit lp nn cc cu trc phn t c hot lc cao v nh
hng phn ho t bo mt cch hiu qu. Trn th gii hin c ba ngn hng d liu gen ln nht l
GenBank (NCBI Entrez Nucleotide - M), EMBL (European Molecular Biology Laboratory - Chu u)
v DDBJ (DNA Data Bank of Japan Nht Bn) lu tr trn 9 t d liu v gen.
Mt s thnh tu ni bt ca tin sinh hc l: chng trnh NMR a chiu thit lp cu trc khng
gian protein. Chng trnh FASTA so snh trnh t gen v protein ra i trc 1990 cho php so snh t
ng, min ph trnh t mt on gen di khong 1000 bp vi trnh t cng b trong vi pht.
Chng trnh BLAST, trung tm NCGR, chip ADN thit lp trc nm 2000 v gn y l n IBM
Blue Gene c bt u, h chng trnh trn gi EMBOSS c bn, chip ADN b gen ngi c
a ra th trng. n thi im ny c trn 60 cng ty ln chuyn dch v v kinh doanh trn lnh vc
sinh tin hc ang hot ng. DNAStar v GCG l hai trong nhng cng ty thnh cng nht trong lnh
vc cung ng phn mm phn tch gen.
1.3.7. Cng ngh nano sinh hc
y l lnh vc a ngnh, tp trung khai thc vt liu, thit b hoc phng php ca cc cht
phm vi kch thc ti hn nm gia chiu di phn t v bc sng nh sng kh kin t 0,1 n 500
nm. Cng ngh nano sinh hc l:
- Phng hng mi cho php thu nhn nhng thng tin v h thng sinh hc mc chm lng t,
to u d nano vi kch thc phn t dng trong chn on bnh.
- Phng php in situ mi cung cp thng tin tt hn v chc nng t bo.
- Cng ngh thao tc ci bin 2 chiu v 3 chiu i vi m v t bo.
- Vn chuyn v phn phi thuc hoc gen vo m v t bo thng qua vic khng ch kch thc
ht, hot ho v gii phng hot cht thuc thng qua c ch v thit b bm kch thc nano, van t bo
v c quan nhn to.
Trong Y hc, ngi ta hy vng phu thut gen, phu thut t bo, tr liu t bo, tr liu gen, tng
hp gen chn on i t bo, cc h thng c kh in t nano y hc - l cc "cng c nano" thng
minh, robot m kch thc nano, s c a vo thc tin trong vi chc nm ti.
CU HI
1. Ai l ngi xc nhn vai tr di truyn ca ADN?
a) Frederick Griffith b) Oswald Avery
c) Hershey v Chase d) Erwin Chargaff
e) Watson v Crick
2. Ai l ngi a ra m hnh xon kp ca ADN?
a) Frederick Griffith b) Oswald Avery
c) Hershey v Chase b) Erwin Chargaff
e) Watson v Crick
3. Bn gen ngi khi hon chnh cho thy c bao nhiu gen m ho cho protein?
a) 10 000 - 15 000 d) 15 000 - 20 000
b) 20 000 - 25 000 c) 25 000 - 30 000
e) 30 000 - 35 000
4. Sinh hc phn t l khoa hc sinh hc nghin cu v
a) Ho hc ca cc phn t sinh hc.
b) nh hng ca cc t bin di truyn.
c) Chc nng ca protein.
d) Chc nng ca gen.
e) Quan h gia gen v sn phm ca n.
5. Ni dung chnh ca hc thuyt trung tm ca Sinh hc phn t:
a) Thng tin khi chuyn sang protein th khng th ly ra li c.
b) Thng tin c lu tr trn ADN c th chuyn sang ARN.
c) Thng tin ch lun chuyn gia cc dng acid nucleic khc nhau.
d) S sao chp, phin m, dch m l cc qu trnh chuyn thng tin trong t bo.
e) Protein khng mang thng tin di truyn.
Bi 2
SAO CHP ADN
MC TIU
- Trnh by c qu trnh sao chp ADN.
- Tm tt c cch thc sao chp ca acid nucleic ca virus.
- M t c qu trnh sa cha ADN.
2.1. KHI NIM
Mt t bo vi khun in hnh c th cha khong 2000 loi protein, cc t bo nhn tht cha
khong 50000. Thng tin v cc loi protein ny c m ho trong cc phn t acid nucleic. a s
loi, vt liu di truyn ch yu l ADN, mt s virus l ARN.
Cc phn t ADN c th khc nhau v thnh phn, trt t sp xp cc nucleotid, nhng u c cu

trc c bn ging nhau. Cu trc xon kp gm hai chui ng - phosphat mt ngoi ca phn t,
cn cc base nit bn trong c ni vi nhau bng cc lin kt hydro (hnh 2.1).
Mc d cc base A, T, G, v C c kch thc khc nhau nhng phn t ADN vn c th t iu

chnh c c cu trc xon kp i xng qua trc. Trnh t ca cc base nit to nn thng tin di
truyn. S khc nhau trong trnh t ny to nn s a dng ca sinh vt. Do , nu xt nghim du n
hng cu th c th bit c 70% b mt di truyn, ngha l hng triu ngi mi c hai ngi ging
nhau, nhng nu xt nghim ADN th bit c 99% b mt di truyn, ngha l trn 70 t ngi mi c
hai ngi ging nhau.
Cc base ni vi nhau bng lin kt hydro theo tng cp b sung. Nh , khi chui xon kp tch
ra cho php cc base c sao chp to nn hai mch mi, c bn ging vi phn t ADN gc v y l
s sao chp c bn trong h thng sinh hc. Cc enzym xc tc qu trnh sao chp ca ADN l cc ADN
- polymerase. Mt s enzym khc cng c vai tr nht nh trong tin trnh ny.
Cc yu t vt l, ho hc c th tc ng ln ADN, lm ph hu ADN hay gy t bin, s h

hi ny c th c sa cha. Cc yu t ny cn c th lm cht t bo hoc lm bin i b gen
(genome). Tt c nhng thay i ny c th di truyn c qua con ng sao chp ADN.
Hnh 2.1. Cu trc phn t xon kp ca ADN
2.2. S SAO CHP CA ADN
Ngi ta a ra hai c ch c bn ca s sao chp ADN: mt c ch bo tn (conservative

mechanism) trong phn t ADN con gm hai chui hon ton mi hoc c ch bn bo tn
(semiconservative mechanism) trong phn t ADN con gm mt chui m kt hp vi mt chui
mi c tng hp. C ch sau c chng minh bng th nghim Meselson v Stahl (1958).
2.2.1. Th nghim ca Meselson v Stahl
Nu t bo Escherichia coli c cung cp mt ngun nit l amoni, chng c th to nn tt c cc

nucleotid thuc nhm purin v pyrimidin cn cho s tng hp ADN. Nu nit c chuyn thnh ng
v nit nng (N15) ADN c to ra s nng hn ADN c tng hp bng cch dng nit nh (N14).
Hai loi ADN nng (N15) hay nh (N14) c th c tch ra bng ly tm vi thang nng cesium
clorid.
Trong th nghim ny, vi khun c cho pht trin vi th h trong mi trng N15 m bo cho
tt c ADN ca vi khun u l loi nng. Sau chuyn t bo vo mi trng c cha N14, nh l
ngun cung cp nit duy nht v cho phn chia trong mi trng N14, khi khi lng ADN tng ln
gp i, ADN c chit ra khi t bo v c ly tm trn thang nng cesium clorid. Nu c ch
bo tn c thc hin, hai bng ADN phi c thy r sau khi ly tm: mt bng nguyn thu (N15) v
mt bng cha N14 (hnh 2.2). Nu theo c ch bn bo tn th ch c mt bng c t trng trung gian
cha N14.5 xut hin. Nu ADN c un nng, ri lm ngui nhanh tch hai mch, ly tm trn thang
nng cesium clorid s thy hai bng tng ng vi ADN N15 v ADN N14.
Hnh 2.2. Th nghim Meselson v Stahl
2.2.2. Cc yu t cn thit cho s sao chp ADN
Tt c mi sinh vt u i hi nhng yu t sau y cho qu trnh sao chp: khun mu (phn t

ADN ban u), M2+, 4 loi desoxyribonucleotid triphosphat (dNTP) v enzym ADN polymerase. Tin
trnh ny c th c tm tt bng phng trnh:
d(NMP)n + dNTP d(NMP)n+1 + PPi
Trong , dNTP l mt desoxyribonucleotid triphosphat v d(NMP)n l mt polymer c n

desoxyribonucleotid.
Vic thm mt desoxyribonucleotid mi vo chui ADN c trnh by hnh 2.3.
Hnh 2.3. S hnh thnh lin kt phosphodiester gia desoxy - ribonucleotid

v 3 - OH ca chui ADN ang sao chp
Pyrophosphat to ra s c thu phn thnh phosphat v c, lm phn ng xy ra theo chiu phi,

ko di chui ADN. C ch tng t cng xy ra trong vic hot ho acid amin, sinh tng hp glycogen
v hot ho acid bo tng hiu sut ca sn phm.
2.2.3. Cc ADN polymerase
ADN polymerase I, II, III c tm thy t bo nhn nguyn thu. c tnh ca cc ADN
polymerase khc nhau c tm tt trong bng 2.1.
ADN polymerase I ca E. coli l mt chui polypeptid ln c khi lng phn t 109000. Trong
mi phn t c cha mt nguyn t km. Enzym polymerase I ch c th xc tc s polymer ho theo
hng 5 3 do s thm mt nucleotid t mt deoxyribonucleotid - 5 - triphosphat vo nhm 3 -
hydroxy ca chui ADN (hnh 2.3).
Tt c ADN polymerase ca t bo nhn nguyn thu cng c hot tnh exonuclease, v chng thu
phn mch n ADN ca chui t u 3 theo hng 3 5 (hot tnh exonuclease 3 5); hoc
chng c th thu phn chui ADN t u 5 (hot tnh exonuclease 5 3). ADN polymerase II ca
E. coli rt ging ADN polymerase I nhng khng c hot tnh exonuclease 5 3. ADN polymerase
III c cu to gm 7 n v nh: , , , , , , v . Tt c 7 n v nh ny u cn cho s sao
chp ca ADN ca E. coli. ADN polymerase III c th l enzym sao chp chnh ca E. coli v cc t
bin vi ADN polymerase III a nhit khng th sao chp c nhit 42oC. ADN polymerase I
chu trch nhim sa cha cc ADN h hng. Vai tr ca ADN polymerase II cha r.
Bng 2.1. c im ca ADN polymerase t E. coli v retrovirus
E. coli
Polymerase Virus
I II III
Khi lng phn t (Mr) 109 000 120 000 180 000 160 000
Cu trc, n v nh (Mr) 1 1 3: 140 000 2: 65 000
25 000 95 000
10 000
Hot tnh polymer ho 5 3 + + + +
Hot tnh exonuclease
5 3 + - + -
3 5 + + + -
t bo nhn tht c 5 loi polymerase c bit l , , , , v (bng 2.2).
Cc enzym ny c c ch tc ng tng t vi cc enzym ca t bo nhn nguyn thu.

Polymerase v lin quan ti s sao chp ca ADN nhim sc th. polymerase cu to t mt chui
polypeptid n, ng vai tr sa cha. Polymerase c tm thy trong ty th v chu trch nhim sao
chp ADN ty th.
ADN polymerase mi c tm thy gn y, c mt s im tng t polymerase v hot tnh.
Bng 2.2. c im ADN polymerase ca nhn tht
Polymerase
Khi lng 300000 45000 140000 180000 - 240000 290000
phn t (Mr)
n v nh 4 1 4 2 2
V tr Nhn Nhn Ty th Nhn Nhn
Hot tnh Sao chp Sa cha Sao chp Sao chp ADN Sao chp
polymer ho ADN nhim ADN ty th nhim sc th ADN
5 3 sc th (si sm) nhim sc
(si mun) th
2.2.4. Qu trnh sao chp ADN E. coli
2.2.4.1. Chc ba sao chp
ADN ca t bo nhn nguyn thu c dng vng, xon kp. Qu trnh sao chp ADN thng bt u
t mt bong bng sao chp, y l ch phnh khi u s tng hp ADN. Cc nt sao chp chnh l
cc chc ba sao chp (hnh 2.4.)
Hnh 2.4. Chc ba sao chp trong qu trnh sao chp ADN
ADN ty th c dng vng v c hai chc ba sao chp. S tng hp ADN ca nhim sc th vi khun
lun lun bt u cng mt im. im ny c gi l v tr Origin hay v tr Ori, l mt vng gm
254 cp base. Dng cc chng vi khun t bin, ngi ta chng minh c rng ch c mt s nh
trong s cc base ny l cn cho s khi u sao chp. E. coli, qu trnh sao chp bt u khi protein
B nhn bit c im khi s sao chp ny (OriC).
Cc si ADN c tch ra v mt si ADN c hng 3' 5' s c sao chp trc tip bi ADN
polymerase III. Si con b sung va to ra c gi l si sm (leading strand). Si gc cn li c
hng 5' 3', khng c sao chp cho n khi mt phn ca si gc c tho xon. Bn sao ny
c gi l si mun (lagging strand). Si mun c tng hp bng mt qu trnh sao chp khng lin
tc. S sinh tng hp si mun v si sm c iu ho bi s to nt tht (loop) ca si mun, nh
si mun cng c tng hp cng hng vi si sm.
Vic sao chp khng lin tc to ra cc on ADN

ngn vo khong 1000 - 2000 nucleotid gi l on
Okazaki (hnh 2.5). Cc on ny sau c ni vi
nhau bi ADN ligase to ra si ADN lin tc.
on mi ARN lun lun cn cho s tng hp ADN.

ADN polymerase i hi mt on ARN ngn c u 3' -
OH t do bt u s tng hp v gn c
desoxyribonucleotid vo mch con, b sung c vi
ADN khun.
Mi ARN c tng hp bi mt enzym c bit c

tn gi l primase. Chng c th ko di s khi ng
chui de novo. Primase nhn ra trnh t c hiu trn
chui ADN n. T bn thn primase khng hot ng
c, n phi to phc hp vi vi chui polypeptid
to thnh primosome chc nng. Cc trnh t ARN c
to ra bi primosome l nhng on ngn khong 5 - 10
base c hiu. Cc protein nhn din c gi l N -
protein, chn cc im (origin) trn ADN, ti primase
c th hot ng. Cc on mi ARN s b ADN
polymerase I loi khi chui ADN. Enzym ny ng thi
lm nhim v lp y cc khong trng bng cc
deoxyribonucleotid thch hp do vic loi mi. Cc on
ADN c ni li vi nhau bi ADN ligase to thnh si
ADN lin tc.
Hnh 2.5. S hnh thnh cc on Okazaki

ca si mun ADN do s sao chp
khng lin tc ca si gc
2.2.4.2. Cu trc theta ( )
S sao chp ADN vng to ra cu trc theta, hnh thnh bi hai chc ba sao chp xut pht t mt v
tr Origin. S tng hp ADN c tin hnh theo c hai chiu thun v ngc kim ng h cng mt
lc. to ra cc si n ADN, hai si ADN gc phi vn xon (quay). C mi 10 base c sao chp
th phn t ADN phi vn xon 1 vng.
Chc ba sao chp ca E. coli cn phi di chuyn vi tc 800 base/giy, i hi ADN gc phi
tho xon vi tc 80 vng/giy. S tho xon ADN gy ra hin tng siu xon (supercoiling). Vic
tho xon si kp dn n siu xon m (xon ngc chiu v lm gim s vng), trong khi , s xon
thm s dn n siu xon dng (xon kp cng chiu, s vng tng) (hnh 2.6).
Hnh 2.6. S hnh thnh cu trc theta do s sao chp hai hng ca ADN kp
C ch sao chp bn bo tn ca ADN i hi cc im ct mt si hay c hai si ADN tch

chui. Mt nhm enzym, gi l topoisomerase s chuyn trng thi topo ca ADN sang trng thi khc.
Topoisomerase II cn gi l gyrase to ra cc dng siu xon m (phi sang tri). Topoisomerase I gn
vi ADN, ct mt si, cho php ADN xon kp quay quanh mt im lm mt i s xon (hnh 2.7 - A).
Phn Tyrosin ca topoisomerase gn vi gc phosphat t do trn ADN gc v phc hp s quay.

Lc by gi, enzym tch khi phc hp v si ADN c to thnh. Loi topoisomerase I tham gia vo
qu trnh to chc ba vi khun cng c tm thy t bo ng vt.
Hai si ADN lng vo nhau c gi l vng lng ghp (catena) hay ADN lng ghp v thng
c to ra khi sao chp ADN vng. Loi topoisomerase II thng lm mt i vng catena, ct si ADN
v phn t ADN si kp i xuyn qua im ct, to phn t ADN kp mi (hnh 2.7 - B).
Hnh 2.7. Vai tr ca topoisomerase

A - Topoisomerase I trong tho xon ADN (tri).
B - Topoisomerase II (Gyrase) trong vic tch hai phn t ADN mch kp lng vo nhau (catena)
2.2.4.3. Enzym helicase
Cc enzym helicase cn thit h tr cho s tho xon ADN gc v hai si n ADN khng th t
nhin tch ra c (hnh 2.4).
Helicase cn ATP nh mt cofactor. Helicase dng nng lng t do khi thu phn ATP i dc
theo si ADN, lm tng tc tch hai si ADN.
C hai loi helicase khc nhau: mt l Rep - protein gn vi ADN gc, trc tip tng hp si sm v
di chuyn theo hng 3' 5'. Helicase th hai gn vi si khun kia tng hp si mun, enzym ny
to phc hp vi primase to mi ARN.
2.2.4.4. Cc protein khc tham gia sao chp
Cc SSB - protein (protein gn mch n) gi cho cc si n ADN do helicase to ra khng chp

li vi nhau. Nh s hin din ca SSB - protein m ADN dng tho xon ri nhau v n nh, l
tng cho vic sao chp. Nu cc SSB - protein b tch ra s lm xut hin cc nt kp tc (hairpin loop)
trong ADN gc v nh vy s ngn chn s sao chp.
Ba protein khc h tr cho vic gn ADN polymerase v hip ng tc ng nh ATPase, thu phn
ATP to nng lng t do cho helicase hot ng. Tc ng hip ng ca cc protein ny trong hnh
2.4 v hnh 2.9 v chc nng ca chng c tm tt trong bng 2.3.
Bng 2.3. Chc nng ca cc protein sao chp quan trng ca E. coli
Protein Chc nng
Protein B Nhn bit im Ori
N - protein Cho php primase hot ng
Primase Tng hp mi ARN
Rep protein Tho xon si sm
Helicase Tho xon si mun
SSB - protein n nh si AND n
ADN polymerase III Tng hp AND
ADN topoisomerase I Ct kha mt si n ADN
ADN topoisomerase II Ct kha c hai si n ADN
ADN ligase Ni cc u ca polynucleotid hnh thnh
2.2.4.5. ADN ligase
ADN ligase gm cc enzym xc tc hnh thnh lin kt phosphodiester gia cc polynucleotid c
hnh thnh.
ADN ligase lin quan ti vic tng hp ADN khng lin tc bng cch ni cc on Okazaki sau khi
thay th cc on mi ARN. Chng cng cn thit cho vic sa cha cc ADN h hng. C ch tc
ng ca cc enzym ny khc vi cc ADN polymerase l do lin kt pyrophosphat ca NAD+ tham gia
(hnh 2.8).
Hnh 2.8. C ch hot ng ca ADN ligase

(NMN = Nicotinamide mononucleotide)
2.2.5. Sao chp ADN t bo nhn tht
2.2.5.1. C ch sao chp
C ch tng hp ADN t bo nhn tht cng tng t nh t bo nhn nguyn thu. im khc
bit ch yu l ADN t bo nhn tht ng cun trong nhiu nhim sc th v di hn. Tc di
chuyn ADN polymerase t bo nhn tht chm hn rt nhiu so vi t bo nhn nguyn thu (khong
50 nucleotid/giy). Nhng t bo nhn tht cha ti 20 000 phn t enzym. Do vy, nhim sc th t
bo nhn tht hnh thnh mt lng ln chc ba sao chp, khong 2000 hay nhiu hn. Cc on
Okazaki nh hn, di khong 40 - 300 base. Do , tc sao chp ADN t bo nhn tht nhanh hn
rt nhiu so vi E. coli. im khc bit c bn l ADN t bo nhn tht c nhiu replicon, v d
Saccharomyces cerevisiae c ti 500 replicon tc l c 500 im Ori.
C ch v s iu ho sao chp t bo nhn tht c nghin cu bng mt m hnh n gin,

l nghin cu s sao chp ADN virus trong t bo ng vt.
Hai loi virus, l Adenovirus v SV40 (virus linh trng) c dng lm cc m hnh ny. C ch
sao chp ca nhim sc th vng ca SV40 cng tng t vi c ch sao chp t mt v tr khi u
(Ori) ca nhim sc th t bo nhn tht.
M hnh to chc ba sao chp t bo nhn tht da trn cc nghin cu ny c trnh by hnh
2.9.
Hnh 2.9. M hnh sao chp t bo nhn tht

(PCNA = proliferating cell nuclear antigen = khng nguyn tng sinh nhn t bo)
C hai loi polymerase tham gia vo qu trnh sao chp: Polymerase tham gia tng hp si sm v
polymerase tng hp si mun. Si mun c tht nt xung quanh polymerase , cho php enzym
di chuyn theo hng chc ba sao chp, ging nh s sao chp t bo nhn nguyn thu.
2.2.5.2. Chu k t bo
S sao chp ADN din ra nh mt phn ca tin trnh phi hp

ca s phn chia t bo. Chu k t bo nhn tht gm 4 pha ring
bit: pha M, G1, S v G2 (hnh 2.10).
Pha G1 l thi k trc khi ADN bt u tng hp, di ca

G1 thay i t pht, gi, tun thm ch n hng nm. T bo khng
bao gi phn chia l t bo trong G1 b ngng tr, thng c
coi nh l giai on Go.
Trong pha S, ADN c sao chp v lng ADN tng gp i

(hnh 2.11). Cc histone mi cng c tng hp trong pha ny, to
thnh 2 b nhim sc th. Tuy nhin, cc nhim sc t vn dnh Hnh 2.10. Chu k t bo nhn tht
chung cho n khi phn chia.
Pha G2 ngn hn pha G1, trong pha ny ngi ta bit c t nhng iu g xy ra t bo. Pha G2
kt thc vi cc du hiu u tin ca s phn bo.
Pha M hay pha phn bo dn n s ho tan mng nhn, s tch nhim sc th v s phn chia t
bo.
Hnh 2.11. Lng ADN c to ra trong chu k t bo
2.2.5.3. S sao chp ca ADN ty th v lp th
Ty th v c khi c lp th, cha ADN polymerase. C th l

ty th c ADN polymerase . S sao chp bt u t vic sao chp
mt si ca ADN gc. S sao chp ny tin hnh cha c na
ng th s sao chp ca si kia bt u. Kt qu to nn cu trc
D. Kt qu ca qu trnh sao chp l hnh thnh s lng ghp ca 2
vng ADN v topoisomerase II cn cho vic tch i 2 vng kp
ADN ty th (hnh 2.12).
2.2.6. S sao chp virus v phage
S sao chp vt liu di truyn ca virus hay phage c thc

hin bnh thng trong trng hp b gen l ADN dng vng i. Hnh 2.12. S hnh thnh cu trc D
Tuy nhin, nu b gen virus l ADN dng thng hay ADN si n trong qu trnh sao chp ADN ty th
hay ARN th cn c c ch sao chp c bit.
2.2.6.1. Sao chp theo kiu dng thng
Nu c dng thng, b gen virus s tr nn ngn hn sau mi ln c sao chp, tr khi virus c c
ch c bit khc phc iu ny. Do ADN polymerase cn c mi ARN v ch hot ng tng hp
ADN theo chiu 5' 3' nn n khng th tng hp chnh xc cc ADN thng (hnh 2.13).
trnh hin tng ny, cc phage s dng mt trong hai cch sau: phage vng ho b gen ca
n nh cc trnh t cos, cn phage T7 thnh lp cc phc ni.
Hnh 2.13. Sao chp ADN theo kiu dng thng
Sao chp ca phage T7
Phage T7 c b gen l dsADN thng, di 39 937 bp. S sao chp bt u ti mt im cch u tn

5 khong 5900 bp v din ra theo c hai hng. Vic sao chp ny gp tr ngi ca s sao chp ADN
thng ni trn. trnh tr ngi ny, b gen ca n c 160 bp u tin bn tri ging ht 160 bp cui
cng bn phi, gi l cc phn tha u. Sau vng sao chp u tin hai phn t con thu c c
mt u dnh vi trnh t ca phn tha (hnh 2.14).
Phn ssADN pha phi (3') ca mt phn t con c th gn vi phn pha tri (cng l 3') ca phn
t khc. Khong trng c lp nucleotid bi ADN polymerase v c ni li bi ADN ligase. Phn t
nh trng thu c c th c ct ra tr li, nhng khi s to ra u dnh 5 thay v 3, u dnh 5 c
th dng lm khun mu hon chnh ADN si i (hnh 2.15).
Hnh 2.14. Bc sao chp u tin phage T7
Hnh 2.15. Phc ni c ct ra v ti to b gen ca phage T7
T7 c gen m ho cho ADN ligase (gen 1.3), SSB (gen 2.5) v ADN polymerase (gen 5) ca ring
n.
2.2.6.2. Sao chp theo kiu ln vng
S sao chp theo kiu theta khng phi l cch duy nht sao chp cc phn t ADN vng. Mt c
ch khc sao chp ADN theo kiu ln vng mc d sn phm to ra khng phi l ADN vng m l
ADN dng thng.
Phn t ADN vng c ct kha mt hay hai lin kt phosphodiester, to ra u 3' OH t do, u
ny c th dng nh khun mu sao chp bi ADN polymerase. Khi u 3' OH c ko di, u 5'
b y ra khi vng di dng si n, trong khi vng ADN ln tip tc ko di u 3. Kiu tng
hp trn vng tng t nh tng hp si sm. Trong lc u 5 b y ra c th dng lm khun mu
tng hp si i theo kiu tng t si mun nu c s hin din ca on mi thch hp (hnh 2.16).
Hnh 2.16. Sao chp theo kiu ln vng
Nu kiu tng hp ny tip tc, s tng hp trn si 5 b y ra s to thnh mt si ADN phc

cha nhiu bn sao lin tip ca phn t ADN vng. Ngha l, nhiu bn sao ca b gen c to ra.
Kiu ny gp phage lambda v to ra nhiu bn sao ni tip, trong khi phage M13 ch to mt
bn sao.
Sao chp ca phage M13
Phage M13 (v cc phage dng si khc) c b gen l mt phn t ADN si n vng. Khi nhim t
bo E. coli, phn t ny c bm vo v c bao bi cc SSB protein. V phage M13 khng c gen
m ho cho ADN polymerase ring nn phi l thuc hon ton vo b my ca t bo ch sao chp.
Mc d b gen l ADN si n ca n rt thch hp lm khun mu tng hp ADN, nhng li khng
thch hp vi vic tng hp ARN bi ARN polymerase hay primase.
Tuy nhin, mt phn ca b gen c dng kp tc si i. Vng ny c th ng vai tr promoter cho

ARN polymerase ca t bo ch phin m mt on mi ARN ngn. S phin m cng lm mt cu
trc kp tc. ADN Polymerase III by gi c th tham gia v tng hp phn t ADN si i, gi l th
sao chp I (RFI replicative form I).
Vic sao chp tip ca RFI khng thc hin theo kiu theta m l ln vng. gp2 endonuclease, c
m ho bi gene 2 ca phage, ct kha ADN ca RFI ti v tr c hiu (im gc trn si dng) v s
sao chp ln vng din ra nhng khng to phn t phc m gp2 endonuclease ct phn t mt ln na
hon tt bn sao. Sn phm thu c gm mt phn t ADN vng n (si b th ra c ni li
thnh vng) v mt phn t ADN si i l RFI. Phn t ssADN vng lc ny c th c sao chp
bng cch lp li t u quy trnh trn.
c c ssADN nh ban u ng gi vo capsid, cc phn t ssADN b th ra cn c bao

bi cc SSB.
Sao chp phage Lambda
Phage lambda c b gen l ADN si i thng. Tuy nhin, mi tn cng ca b gen l mt on

si n gi l u dnh, hai on hai u c th gn b sung vi nhau. Chng di 12 nucleotid v c
gi l v tr cos.
Sau khi phage nhim t bo E. coli v tim ADN vo, nhim sc th ca n vng ho nh cc v tr
cos. Nh n trnh b ct bi cc exonuclease ca vi khun v c th c sao chp theo chu trnh tiu
gii tim n. Qu trnh sao chp ca phage lambda gm hai giai on.
Giai on sm
Phage lambda khi u sao chp theo m hnh theta. im gc sao chp (ori) nm gen O, c sn
phm tng t DnaA. Phn t ny gn trnh t lp im gc v khi u vic tch hai si ADN ti
ch. Mt gen khc, gen P, c sn phm vi chc nng tng t DnaC. Sn phm ny gip DnaB gn
vo ADN tch mch. Sau , cc thnh phn khc ca b my sao chp ca vi khun gn vo v bt
u vic sao chp. Kiu sao chp ny ko di khong 5 - 15 pht.
Giai on mun
Sau 15 pht, phage lambda chuyn sang kiu sao chp ln vng. Hin cha bit nguyn nhn v c
ch ca s chuyn i ny.
Hu qu ca c ch ln vng l ADN ca phage c sao chp thnh mt ADN kp di cha nhiu

bn sao lin tip ca b gen phage, cch nhau bng cc trnh t cos. Phage c gen m ho cho mt
enzym l Terminase, enzym ny nhn din v tr cos ( dng si kp) v ct ti bin phn t ADN
di thnh cc bn sao n ca b gen phage vi cc v tr cos hai u.
In vivo, qu trnh x l bn sao ln thnh b gen n c s tham gia ca cc protein capsid. Cc

protein ny nhn din v tr cos th nht v nu khong cch n v tr cos th hai phi nm trong gii
hn 75 - 105% so vi chiu di nguyn thu ca b gen phage th capsid s ng gi ADN to thnh
phage hon chnh.
2.3. SA SAI TRONG SAO CHP V KHI KHNG SAO CHP
ADN c sao chp bi ADN polymerase vi chnh xc cao (c khong 108 - 1012 base th c
mt base sai c gn vo mi ln sao chp). S sao chp chnh xc ca phn t ADN c tm quan
trng sng cn i vi hot ng bnh thng ca t bo.
2.3.1. Sa sai trong sao chp
Ngi ta dng cc nucleotid v ADN polymerase tng hp ADN in vitro. Sai st trong trng
hp ny l 105 tc l trong 100 000 nucleotid c mt sai st. E. coli c 3.106 cp base, vy mi ln
sao chp theo tnh ton nh trong thc nghim phi c ti 30 sai st. Tuy nhin, trn thc t sai st trong
t nhin thp hn nhiu.
Bng cch nh gi tn s cc t bin mi xut hin trong qun th ln v theo di bin i enzym
no trong nui cy m t bo, ngi ta tnh c sai st trong c th sinh vt khi sao chp in vivo l
109 tc l mt sai st trn mt t base. Nh vy t bo c 3.109 cp base mi ln sao chp ch c 3 sai
st.
Mc chnh xc cao ca sao chp trong c th sinh vt c c nh cc c ch sa sai:
Hng sao chp bao gi cng t u 5 3 vic sa sai chnh xc.
t bo nhn nguyn thu ADN polymerase I v III va polymer ho, va c hot tnh exonuclease
5 3 v 3 5. Nu trn ng di chuyn polymer ho gp base sai, ADN - polymerase s li
li ct b theo hng 3 5 (hot tnh 3 5).
t bo nhn tht, hot tnh exonuclease ch c c polymerase v . Khng tm thy hot

tnh exonuclease polymerase , c th do mt tiu n v ca n b mt i trong qu trnh chit tch
enzym ny.
ADN ty th d b t bin, nhng polymerase ca n li khng c hot tnh exonuclease.
2.3.2. Sa sai khi khng sao chp
Phn t ADN c th b bin i ngay c khi khng sao chp. Cc bin i t bin ny xy ra vi
tn s kh cao. Nh c ch sa sai nn tn s t bin c duy tr mc thp.
Cc enzym nhn bit gn vo cc trnh t sai v ct ri on sai, ri dng mch n ng lm khun

mu tng hp li ch b sai cho ng.
Hng lot enzym c hiu lm nhim v d tm v sa sai. C khong 20 enzym r sot dc ADN
d tm cc base b bin i ho hc, mi enzym c mt chc nng chuyn bit cho mt loi sai hng.
Khong 5 enzym khc c hiu cho cc lin kt cng ho tr sai gia base vi cc cht ho hc khc
hoc gia cc base k nhau trn mt mch. Mt s enzym khc pht hin s bt cp sai, nh trng hp
mt purin. Tng cng c khong 50 enzym chuyn bit pht hin v sa cc sai hng trn ADN.
Tm tt
S sao chp chnh xc ca ADN nhim sc th l mt s khi u cn thit cho s sinh sn. S sao
chp theo c ch bn bo tn v mi phn t ADN mi bao gm mt mch ADN gc v mt mch mi
c tng hp. S tng hp ADN c thc hin bi cc ADN polymerase chuyn bit, chng dng
ADN c sn lm khun mu. Ch c loi ADN polymerase III l cn thit cho s sao chp ADN ca t
bo nhn nguyn thu. Tuy nhin, cc t bo nhn tht c mt s polymerase tham gia vo qu trnh
sao chp. Mt s protein khc cng tham gia vo qu trnh tng hp ADN c t bo nhn tht ln t
bo nhn nguyn thu.
Cc phn t ADN c th b h hi bi cc tc nhn khc nhau v s h hi ny c th c sa cha

hn ch cc t bin c hi. Qu trnh sa cha c iu khin bi cc h thng c s tham gia ca
enzym.
CU HI
1. ADN sao chp theo c ch bn bo ton v t mt gen ban u to ra
a) 2 gen con cha cc nucleotid c v mi xen k.
b) 2 mch n ADN cha cc nucleotid c v mi xen k.
c) 1 gen con hon ton mi, 1 gen con hon ton c.
d) 2 gen con, mi gen cha mt mch mi, mt mch c.
e) 1 gen ban u ng thi tn ti vi 2 gen con va mi va c.
2. Chn t hp sai
a) ADN polymerase Nhn Sao chp si mun.
b) ADN polymerase Nhn Sao chp si sm.
c) ADN polymerase Ty th Sao chp ADN.
d) ADN polymerase Nhn Sao chp si sm.
e) ADN polymerase Nhn Sao chp ADN.
3. ADN polymerase ng vai tr sa cha ca t bo nhn tht
a) I b) II
c) d)
e) b v d
4. Bong bng sao chp cn c gi l
a) Nt sao chp b) Chc ba sao chp
c) V tr Origin d) V tr Okazaki
e) Tt c u sai
5. no ng vi on Okazaki t bo nhn nguyn thu?
a) Gm khong 1000 - 2000 nucleotid
b) c ni li bng ADN ligase
c) Ni vi nhau to thnh si sm
d) Cn gi l loop
e) a v b
6. V tr Origin
a) im khi u s sao chp b) c nhn din bi protein B
c) Gm 254 cp base d) a v b
e) a, b v c
7. Primase l enzym
a) T bn thn khng hot ng c
b) Gm nhiu N - protein gn
c) Cn gi l primosome
d) Lp y cc GAP bng dNTP
e) a v c
8. Primase bt u hot ng khi
a) N - protein c nhn din
b) N - protein nhn din c Ori
c) Protein - B nhn din c Ori
d) To phc hp vi cc chui polypeptid
e) To phc hp vi N - protein
9. Bn sao ADN vng mi c tng hp, tch ra khi bn gc nh
a) Topoisomerase I b) Topoisomerase II
c) Tho xon m d) Phn Tyrosin ca Topoisomerase
e) Gc phosphat t do ca Topoisomerase
10. Phage lambda sao chp b gen ca n theo kiu
a) Theta b) Theta v ln vng
c) Sao chp ADN thng d) Sao chp ADN vng
e) Sao chp ngc
Bi 3
CC LOI ARN
MC TIU
- M t c cu trc ca tng loi ARN.
- Gii thch c chc nng v vai tr ca cc loi ARN.
- Trnh by c cc cch thc bin i cc bn sao nguyn thu ARN v cc chc nng ca s
bin i ny.
3.1. KHI NIM
ADN mang thng tin di truyn ca t bo nhng bn thn n khng trc tip ch huy qu trnh tng
hp protein, v ADN nhim sc th ch mang mt bn sao duy nht cho mi gen, trong khi t bo c
hng nghn phn t protein cng loi tn ti. Mt khc, khng phi tt c cc gen u c biu hin
thnh protein cng lc m cc gen khc nhau s biu hin khc nhau ti mi thi im trong vng i
ca t bo v tu theo iu kin mi trng. Do vy cn c c ch trung gian khuch i thng tin di
truyn t ADN n protein, ng thi kim sot s biu hin ca gen theo nhu cu ca t bo. ARN
ng vai tr trung gian quan trng ny. T mi gen c th c nhiu bn sao ARN ch huy qu trnh
tng hp protein, ng thi vng i ca ARN ngn nn s biu hin ca gen qua trung gian ARN d
dng c kim sot bi nhu cu ca t bo v iu kin mi trng.
Ngoi vai tr trung gian trong qu trnh biu hin gen, nhiu ARN cn c vai tr cu trc hay xc
tc.
t bo nhn tht, ARN c tng hp ngay trong nhn (cha ADN), sau i vo t bo cht
tham gia tng hp protein. Nhng t bo giu ARN tng hp protein nhiu hn. V d: cc t bo tng
hp nhiu protein nh gan, l lch, tuyn t ca tm cha ARN nhiu hn so vi t bo t tng hp
protein nh thn, tim, phi.
V cu trc, ARN ging ADN: mch polyribonucleotid thng ca ARN cng cha 4 loi
ribonucleotid: Adenin (A), Guanin (G), Cytosin (C) v Uracil (U) thay cho Thymin (T).
Tt c ARN u c tng hp t cc gen tng ng trn ADN v u bao gm:
- Mch n polyribonucleotid
- ng ribose (5C)
- Cc base nit Adenin, Guanin, Cytosin v Uracil ( ADN l Thymin).
ARN gm cc loi:
- mARN - ARN thng tin.
- rARN - ARN ribosom.
- tARN - ARN vn chuyn.
- pre - rARN (tin rARN) l ARN c tng hp t ADN, sau khi chng c ct ni s tr thnh
rARN.
- pre - tARN (tin tARN) l ARN tng hp t ADN, sau khi c ct ni s tr thnh tARN.
- hnARN - ARN nhn khng ng nht, s to mARN sau khi c ct ni.
- snARN - ARN nhn nh.
- scARN - ARN t bo cht nh.
ARN c th dng t do hoc gn vi protein thnh cc phc hp nucleoprotein gi nhiu vai tr

quan trng trong hot ng sng ca t bo.
C ti 7 loi ARN nh khc nhau, trong c mt s snARN c lin quan n vic x l hnARN
thnh mARN chc nng. Gn y ngi ta mi tm thy mt vi loi ARN c c t bo nhn nguyn
thu v t bo nhn tht l thnh phn ca enzym c chc nng xc tc trc tip.
3.2. CC ARN V VAI TR CA CHNG
3.2.1. ARN ribosom
rARN l thnh phn cu to ribosom, chim phn na khi lng ca ribosom.
Trong dch nui cy Escherichia coli ang giai on phn chia nhanh, ribosom chim ti 1/3 khi
lng t bo. Khong 1/2 khi lng ribosom l ARN nn rARN chim n 75% ca tng s ARN.
Mi mt ribosom c Mr = 2.5 - 4.5 106. C 3 loi ribosom khc nhau, thng c phn bit da
vo h s lng ca chng khi siu ly tm. Ribosom ca Eubacteria v lc lp c ch s lng khong 70S.
Ribosom ca t bo nhn tht l 80S, ca ty th ng vt c v khong 50S.
Tt c ribosom u c cu to bi 2 tiu n v (subunit) c chc nng v cu trc ring bit, cc

tiu n v ny c th tch ra v gn li vi nhau mt cch thun nghch tu theo nng ca Mg2+
(hnh 3.1).
Hnh 3.1. Thnh phn ca ribosom 70S (t bo nhn nguyn thu) v 80S (t bo nhn tht)
Cc ribosom ca lc lp v ca t bo nhn nguyn thu c h s lng khi ly tm l 70S gm 2 tiu

n v:
- n v ln 50S cha 1 rARN 23S v 1 rARN 5S v 34 protein.
- n v nh 30S ch cha 1 rARN 16S v 21 - 23 protein.
Cc ribosom ca t bo nhn tht c h s lng khi ly tm l 80S gm 2 tiu n v:
- n v ln 60S cha 3 loi rARN khc nhau: 1 rARN 28S, 1 rARN 5,8S, 1 rARN 5S v 45
protein.
- n v nh 40S ch c 1 rARN 18S v 33 protein.
3.2.1.1. Ribosom E. coli
Ribosom E. coli c nghin cu y , cc chui polynucleotid ca 3 loi rARN cng vi cu

trc bc 1 ca 55 protein ca chng c khm ph.
Cc protein ribosom (r - protein) ca n v nh 30S c t tn t S1 - S21. Cc protein ca n v

ln 50S c t tn t L1 - L34. Hu ht cc r - protein c khi lng tng i nh (trung bnh Mr 17
000) v cha nhiu acid amin base.
Hu ht r - protein ca ribosom E.coli c cc trnh t bc 1 khc nhau, ngoi tr L7, L12 v L8. Cc
protein L7 v L12 c nhng trnh t c xc nh, L7 l mt dng aminoacetyl ho L12 (s acetyl ho
cn thit cho hot tnh GTPase trong qu trnh dch m). L8 l phc hp ca mt phn t L10 lin kt vi
L7 v L12. i khi trong qu trnh tinh ch, phc hp ny khng tch c.
3.2.1.2. S lp rp ribosom
Cc thng tin cho vic t lp rp ca ribosom c cha ng trong cc phn t thnh phn rARN
v protein. Cc tiu n v ribosom c th ti hp li in vitro c s dng rng ri cho vic nghin
cu qu trnh lp rp ca ribosom. Cc tiu n v c treo trong mt dung dch c nng mui cao
(v d LiCl 4 M v urea 8 M) ngn cn i lc yu khng ho tr gia protein v cc phn t ARN, lm
cho cc protein tr nn khng bn vng vi ribosom. ARN v cc protein khng lin kt c tch ra
khi nhau. Chng c thm phn loi b mui v urea, sau vi nhau. Di nhng iu kin
thch hp, cc tiu n v c t hp li theo chc nng ca chng v khng th phn bit c vi s
t hp cc tiu n v t nhin.
Khi dng s t hp khc nhau gia cc thnh phn ring l ca ribosom, ngi ta c th xc nh
c cc trnh t lin kt trong qu trnh lp rp.
V d, 6 r - protein ca 30S ca E. coli c v tr kt ni c lp v chuyn bit vi rARN 16S.
3.2.1.3. Cu trc ca rARN
Nh tin b ca cng ngh di truyn, ngi ta c th to dng v xc nh trnh t ca cc rARN t

sinh vt khc nhau, t ty th v lp th. Mc d chng rt khc nhau v kch thc (bng 3.1) nhng u
c th xc lp c bn cu trc bc 2.
Bng 3.1. Kch thc theo nucleotid ca cc rARN khc nhau
Ngun gc ribosom rARN ca tiu n v ln rARN ca tiu n v nh
T bo cht chut 4950 1859
T bo cht nm men
3392 1787
(Saccharomyces cerevisae)
Escherichia coli 2904 1541
Lp th thuc l 2950 1846
Ty th linh trng 1559 854
Cu trc bc 2 chnh xc ca rARN c tm quan trng trong vic lp rp ribosom v chng c lu

tr tt trong qu trnh tin ho. Tt c cc protein ca E. coli c gn kt vo rARN cng c cu hnh
bc hai nh vy v nu rARN b bin tnh th protein khng gn kt c.
Mt c tnh trong cu trc bc 1 ca rARN l s methyl ho nucleotid. S methyl ho ny xy ra

sau khi phin m v c thc hin bi ARN - methylase trong c s dng S - Adenosyl Methionin
(SAM) nh l c cht cung cp methyl. rARN ca t bo nhn nguyn thu v ca cc bo quan thng
methyl ho base khi u (5 - methylcytosin), cn rARN ca t bo cht t bo nhn tht thng
methyl ho v tr 2 ca ng ribose (2 - 0 - methyladenosyl). Cc v tr ca phn methyl ho trong
chui rARN c lu tr tt trong cc loi khc nhau. Ngi ta cho rng s methyl ho ny ng vai tr
bin i bn sao s cp ca rARN.
3.2.1.4. S bin i ca rARN
Hu nh tt c cc c quan, nhng rARN ln ca 2 tiu n v ribosom u c to thnh t s

bin i v x l ca cc ARN ln hn, l cc pre - rARN. S sa i xy ra trong nhn t bo, m
bo cho 2 ARN ln c to ra cc ARN trng thnh c khi lng tng ng vi tng tiu n v.
Cch bin i ca rARN t bo HeLa (ngi) c minh ha hnh 3.2.
Hnh 3.2. Qu trnh sinh tng hp cc tiu n v ribosom cc t bo HeLa
Pre - rARN 45S (gm khong 12000 nucleotid) l bn sao ca cc gen ARN - ribosom. Cc pre -
ARN c vi trm bn sao v tp hp li to thnh phn u trong mt vng nhim sc th v c xem
nh vng thit lp nhn. rARN 45S c bin i do a vo khong 110 nhm methyl cc nucleotid
c hiu. Hu ht nhng phn c methyl ho u c bo tn rARN trng thnh. Hng lot din
bin trong v ngoi nhn to ra c cc tin th trung gian 32S v 20S t pre - rARN 45S. Sau ,
pre rARN 32S ct ni thnh 28S v 5,8S, cn 20S thnh 18S.
Qu trnh sa i ca pre - ARN khc nhau v chi tit gia t bo nhn tht v t bo nhn nguyn
thu. t bo nhn nguyn thu, chui rARN 5S l thnh phn ca n v phin m ca c rARN 16S
v rARN 23S. Hn na, gen rARN t bo nhn nguyn thu li c cha 2 chui tARN hay nhiu hn.
Nhng s phn ct u tin ca pre - rARN c xc tc bi ARNse III c hiu cho ARN mch kp
(hnh 3.3).
Hnh 3.3. Qu trnh ct ni ca pre - rARN E. coli di khong 5000 nucleotid.
3.2.2. Cc ARN vn chuyn
3.2.2.1. Cu trc
Nm 1950, F. Crick nu ra gi thit v cht ni. ng cho rng trc khi gn thnh polypeptid, cc
amino acid phi gn qua cht ni trung gian, cht ny s bt cp c hiu vi cc base trn mARN. Vo
nm 1957, M. Hoagland v cc cng s tm ra tARN v chng minh c rng mi phn t tARN
gn vi mt phn t amino acid v mang chng n ribosom.
t nht, mi loi tARN c hiu cho mt loi amino acid. V d: Arginine ch gn c hiu vi ARN
vn chuyn arginine. Tuy nhin, tt c cc tARN c mt s c tnh cu trc chung: chiu di khong 73
- 93 nucleotid, cu trc gm mt mch cun li nh hnh l ch ba (hnh 3.4). u mt 3 c trnh t kt
thc CCA.
Hnh 3.4. Cu trc alanin - tARN

Cu trc hnh l ch ba ca tARN - Alanin ca nm men cho thy trnh t nucleotid
hon chnh. th, mt vi tARN thiu vng xon dihydrouracil.
Cc nucleotid thng gp: I = Inosine
T = Ribothymidine
= pseudouridine
m1G = methyl guanosine
m22G = dimethylguanosine
m1I = methylinosine
D = dihydrouridine
Cc enzym c hiu l aminoacyl tARN synthetase gn mi acid amin vi tARN tng ng nh

nng lng ATP to ra aminoacyl - tARN. Phc hp ny n ribosom gn vi mARN. tARN gn vi
mARN bng s bt cp b sung nh i m.
3.2.2.2. Bin i tARN
t bo nhn nguyn thu v t bo nhn tht, cc tARN u c tch ra t nhng phn t ban
u di hn nh mt s phn ng x l. E. coli, cc gen ca tARN thng c pht hin trong cc
operon hn hp, cng lc m ho cho 4 tARN (tARNTyr, tARNThr, tARNGly, tARNThr) v mARN
cho s tng hp protein - yu t ni di Tu.
Qu trnh ct ni thu gii u tin ca bn sao s cp c xc tc bi RNse, to cc u tn 5 ca

phn t tARN.
RNse P l mt enzym, gm 1 ARN c 375 nucleotid v mt protein c trng lng phn t 20 000.
Trong cc iu kin sinh l, c 2 thnh phn protein v ARN u cn nhng yu t hot ng, nhng
khi nng Mg++ cao th ch c thnh phn ARN c hot tnh xc tc. Theo sau hot ng ca RNse P,
mt s phn ng trong nhn v bin i base s to nn tARN hon chnh.
E. coli, tARN c trnh t cui l CCA - OH, phin m t cc base b sung c trong ADN ca
nhim sc th. Trong khi t bo nhn tht nhng base ny c thm vo sau khi phin m (hnh
3.5).
3.2.2.3. Cc phn ng trong qu trnh x l tARN
Mt s phn ng ch yu ca ARN vn chuyn trong qu trnh sinh tng hp protein:
- Aminoacyl ho
- Formyl ho tARN m u (t bo nhn nguyn thu)
- Gn nhng yu t ni di
- Gn ribosom
- Nhn din codon - anticodon.
Hnh 3.5. Qu trnh bin i ca tARNTyr nm men
tARN c nhiu im nhn din enzym. Mt s im nhn din c th cho tng tARN ring bit hay
cho mt s loi ng ng (nh aminoacyl ho, formyl ho, nhn din codon - anticodon). Trong khi ,
c nhng im nhn din c hiu cho tt c tARN ( l, s tng tc vi nhng yu t ni di I, II v
s gn kt ribosom).
3.2.3. ARN thng tin
mARN c tng hp khi enzym ARN polymerase II tip xc vi trnh t promoter trn phn t
ADN. mARN nguyn vn ca t bo vi khun v ca t bo nhn tht cha trnh t nucleotid nhiu hn
s c dng m ho protein.
3.2.3.1. Cu trc
Phn t mARN gm 3 phn:
1. on 5 khng m ho
2. Vng m ho
3. on 3 khng m ho
ADN polymerase khi u s phin m mt on nm ngay trc vng m ho cho phn t

protein c gi l on 5 khng m ho. Do , mARN c on u mang cc tn hiu cho ribosom
nhn bit gn vo khi dch m.
ui 3, sau du kt thc c on 3 khng m ho l ni gn poly A (t bo nhn tht).
Cc mARN ca t bo nhn nguyn thu c thi gian bn hu ngn: trung bnh 2 pht, ca t bo
nhn tht t 30 pht n 24 gi.
Hnh 3.6. Intron v exon t bo nhn tht
on 5 v 3 l trnh t khi u v kt thc khng c dch m, do vy mARN nguyn vn ca t

bo vi khun v t bo nhn tht cha trnh t nucleotid nhiu hn s c dng m ho protein. Cc
trnh t ny c phin m t si ADN, k hiu 5UTR v 3UTR. Tuy nhin, do i lc khng n nh
vi enzym RNase, on 5 hay 3 c th lm gia tng hay gim n nh ca phn t ARN. Cc on
UTR gip phn t ARN tn ti lu hn trong t bo trc khi b thoi ho, do vy s tng hp c
nhiu protein hn. Nu khng c cc on UTR, ARN s b thoi ho nhanh hn v lng protein sn
xut s t hn. Hn na, trnh t UTR c th thc y qu trnh dch m hiu qu hn.
3.2.3.2. mARN t bo nhn tht
t bo nhn tht, sau khi mARN c tng hp ti nhn, chng phi tri qua 2 giai on "hu
phin m":
1. Giai on gn chp (capping): u tin, nhm triphosphat ti u 5 ca phn t ARN mi tng

hp s b ct nh enzym phosphohydrolase ct lin kt - phosphodiester, loi b v phosphat. Tip
theo, enzym guanyltransferase gn guanin v phosphat vo phosphat ca u 5 bng kin kt 5 - 5
triphosphat. Sau , v tr N7 ca guanin s b methyl ho do enzym guanin - 7 - methyltranferase. Cui
cng, enzym 2 - O - methyltransferase s methyl ho v tr 2 ca ng ribose.
Phn ng methyl ho chp c th xy ra theo 1 trong 3 cch:
a) Chp 0: nhm methyl gn vo v tr G7 do enzym guanin - 7 - methyl transferase xc tc. Phn

ng ny xy ra tt c mARN t bo nhn tht.
b) Chp 1: nhm methyl gn vo v tr 2 - OH ng ribose ca nucleotid u tin. Phn ng do

enzym 2 - O - methyl transferase xc tc, xy ra hu ht mARN t bo nhn tht.
c) Chp 2: phn ng methyl ho tng t xy ra nucleotid th 2 vi t l 10 - 15%.
Hnh 3.7. Gn chp mARN ca t bo nhn tht

1st base: base cacbon v tr s 1; 2nd base: base cacbon v tr s 2; 5capping:
gn chp u 5; 5 - 5 triphosphaste linkage: lin kt triphosphat 5 - 5.
Ngoi ra, cn c qu trnh methyl ho ni phn t xy ra ti v tr N6 ca adenin, vi tn sut 0,1%.
Giai on ny bo v phn t ARN khng b ct lin kt phosphoester do cc phn t hydro ln cn

v chp 5 l du hiu nhn bit v tr gn ca mARN trn ribosom khi dch m.
2. Giai on gn ui poly A vo u 3: ng vt c v khong 50 - 250, cn nm men

khong 100 nucleotid Adenin c gn vo u 3. ui poly A gn vo pre - mARN c trnh t nhn
din AAUAAA. Qu trnh ny do enzym poly A polymerase xc tc. Chc nng ui poly A tng t
chp 5 trong vic bo v phn t ARN v gn vi ribosom. Do vy, thi gian bn hu c ko di v
lng protein tng hp cng nhiu hn. Ngoi ra u 3 cng c vai tr trong vic vn chuyn mARN t
nhn ra t bo cht.
Hnh 3.8. Cu trc mARNca t bo nhn tht
3.2.3.3. mARN ngc
mARN ngc c th ngn cn qu trnh dch m nhiu t bo nhn tht khi chng c trnh t b
sung vi trnh t ca mARN. iu ny c ngha l mt gen khng biu hin c thnh protein nu
trong t bo c mARN ngc. y c th l mt c ch chng li hin tng gen nhy ngc hay virus;
do c 2 trng hp u dng si i mARN lm trung gian. Trong sinh ho, hin tng ny c dng
nghin cu chc nng ca gen bng cch thm vo mARN ngc.
3.2.4. Cc ARN nhn nh v cc ARN t bo cht nh
T bo nhn tht c cha nhiu phn t ARN nh khc ngoi tARN v 5S v 5,8S rARN. In vivo,
nhng phn t nh ny phc hp vi cc protein c hiu to nn cc ht ribonucleoprotein (RNP) c
gi l snRNP hoc scRNP tng ng cho nhn hay cho t bo cht. Chng di khong 90 - 300
nucleotid.
Vic bin i tin ARN thnh ARN c thc hin vi s tham gia ca nhng phn t nh l cc
spliceosom, chnh l cc snRNP. Ngy nay tm thy c 7 loi snRNP t U1 n U7. U1 v U2 c vai
tr trong vic sa i hnARN thnh mARN hon chnh, cn U3 c vai tr trong sa i pre - rARN
thnh rARN t bo cht. Cc ARN c kh nng xc tc c gi l ribozyme. Ngi b bnh t min
hng ban Lupus (systemic lupus erythematosus), do bnh nhn to ra khng th gn vi mt trong cc
protein U1 - U7 snRNP.
scRNP c vai tr bit r nht l 7SL ARN, mt thnh phn ca phc hp nhn din tn hiu xut
SRP. SRP cu to bi mt scARN c cha 294 nucleotid v 6 polypeptid c Mr t 9000 - 72000. Khi
mARN ca protein xut bt u c dch m, SRP tng tc c hiu vi on peptid tn hiu xut ca
protein mi sinh v vi ribosom, ngn chn s dch m cho n khi ribosom tip xc vi li ni cht,
khi n ri khi ribosom v tn hiu xut ca protein mi sinh gn vo b my chuyn v protein trn
mng li ni cht v s tng hp protein c tip tc, protein sinh ra c chuyn vo trong li ni
cht.
Hnh 3.9 l s m t phn ng t ct ni, trong trnh t intron t xc tc qu trnh t ct ri

khi phn t rARN loi Tetrahymena. Phn ng c bt u khi nucleotid G gn vo intron, ng
thi ct mch ARN, u 3 ca ARN mi va to thnh gn vo u bn kia ca intron hon thnh

phn ng ni lin.
Ch c mt t l nh cc ADN sinh vt t bo nhn tht m ho protein. Cc ADN d tha nh vy

c pht hin gia cc gen cu trc. Trong s nhng ngi u tin pht hin cc on ADN d tha
ny (intron) c Pierre Champon v ng s i hc Strasbourg (Php) qua nghin cu s iu ho di
truyn trong tng hp ovalbumin lng trng trng g. Ngi ta thy trong nhn t bo ch c nhng
phn t ln hn mARN hon chnh trong t bo cht rt nhiu v gi chng l nhng ARN nhn khng
ng nht, hnARN. So snh ADN ca gen v ADN phng x c tng hp t sao chp ngc ca
mARN trong t bo cht, Champon nhn xt rng cc on nucleotid di c mt trong ADN ca gen ban
u khng thy trong cADN phng x ca mARN t bo cht. Cc on di ny khng dch m, xut
hin khng ch on u hoc cui ca gen m c nhng on gia, lm gin on cc on m ho
protein (exon). Gen m ho ovalbumin b gin on bi 7 intron.
Hnh 3.9. Phn ng t ct ni (self - splicing)

ca rARN loi Tetrahymena
3.2.5. Tm tt cc qu trnh ct ni ARN t bo nhn tht
3.2.5.1. Khi nim
B gen t bo nhn nguyn thu l b gen gin on. Trnh t khng m ho xen k cc trnh t m
ho. Qu trnh loi b cc trnh t khng m ho gi l qu trnh ct ni ARN.
Exon l trnh t biu hin, c mARN hon chnh. Tuy nhin, u tn cng 5 v 3 c th khng
dch m thnh trnh t protein, do vy trnh t exon khng hon ton l trnh t m ho.
Intron l trnh t thng khng cha gen, xen k cc exon, hin din hu ht cc phn t ADN ca
t bo nhn tht bc cao. Chng thng c kch thc ln hn cc exon. Intron c rt t t bo nhn
tht bc thp, nm v hon ton khng c vi khun tr mt vi ngoi l.
3.2.5.2. Ct ni ARN
Qu trnh ct ni xy ra cc phc hp bn sao nguyn thu pre - ARN hay hnARN vi protein
nhn gi l hnRNP. Qu trnh ny din ra trong nhn t bo trc khi di chuyn vo t bo cht tham
gia dch m.
Nguyn tc GU - AG
- GU lun tn cng u 5 ca intron: v tr cho.
- AG lun tn cng u 3 ca intron: v tr nhn.
Bt k v tr cho no cng c th gn vi v tr nhn.
- C ch ct ni thng thng: 2 phn ng chuyn v ester ho.
- Phn ng chuyn v ester ho I: nhm 5' - OH a nhn ca exon gn vo v tr 5P ca intron.
- Phn ng chuyn v ester ho II: nhm 3 - OH a nhn ca exon gn vo v tr 3P ca intron
- Ni cc exon, loi b intron.
Mi bc l mt phn ng chuyn v ester ho, lin kt phosphodiester mi c to thnh. Phn

ng ny khng cn cung cp nng lng.
Ba h thng ct ni c bn xy ra theo cc c ch chnh:
1. T ct ni
ARN ng vai tr enzym, gi l ribozym. Loi ny c 2 nhm intron: nhm I v nhm II
- Nhm intron I: t ct ni vng 9 ca cu trc bc 2. Qu trnh ny xy ra gen rARN ca t bo

nhn tht bc thp v nm.
- Nhm intron II: tng t c ch ct ni bng spliceosom nhng c xc tc bi cc intron ARN.
2. Intron c loi b khi tin tARN nhn ca nm men tng t qu trnh x l tin tARN.
3. Ct ni nh spliceosom
Spliceosom l phc hp cha nhiu loi ribonucleopotein. Chng nhn din cc trnh t exon
intron loi b intron v ni cc exon. Qu trnh ny thng xy ra ARN nhn ca t bo nhn tht
bc cao
Hnh 3.10. Cc protein tham gia tng hp v to mARN hon chnh
3.2.5.3. Ct ni mARN t bo nhn tht
Spliceosom
Spliceosom cu to bi khong 40 protein quy nh ct ni v cc snARN. Cc phc hp

ribonucleoprotein ny c gi l snRNP hay snURP. Cc snRNP gm c U1, U2, U5 v U4/U6 cha
snARN U1, U2, U4, U5, U6. Cu trc ny n nh cc t bo nhn tht.
Phn ng ct ni: gm 3 giai on chnh
1. Giai on 1: v tr cho b ct, u 5 gn vo v tr nhnh ca intron bng lin kt 5 - 2

phosphodiester to si i tm thi. V tr nhnh gm khong 30 nucleotid nhnh ln ca v tr nhn,
c trnh t tng ng.
2. Giai on 2: v tr nhn b ct, cc exon c ni li, intron b loi b khi si i.
3. Giai on 3: si i b "b nhnh" v thoi ho.
Vai tr ca snRNP trong 3 giai on:
- snRNP U1 gn vo v tr cho.
- snRNP U2 to lin kt hydro vi v tr nhnh ca intron. Giai on ny s dng ATP thu phn v
"yu t tr gip" snRNP (U2AF).
- SnRNP U1 tng tc vi snRNP U2, gip v tr cho v v tr nhn tin li gn nhau.
- Phc hp U5/U4/U6 gn vi U1 v U2 s dng ATP thu phn; U5 gn vi v tr cho ca intron;

U6 gn vi U2: y l spliceosom hon chnh.
ATP cung cp nng lng cho mi giai on. U4 tng tc vi U6 bng lin kt hydro.
- U5 di chuyn t phn intron sang phn exon ca v tr cho, U2 thay th U4 lin kt vi U6 nh lin
kt hidro.
Cu trc bc 2 ca snARN U khng thay i. Gc vung l trnh t lin kt vi snRNP protein: a)

Vng 5 ca snARN U1 to cp base vi nucleotid ti u 5 ca intron v u 3 ca 5exon ca pre -
mARN; snARN U2 to cp base vi trnh t c im nhnh A. b) Mt vng ca snARN U6 to cp base
vi u 5 ca snARN U4. c) Trong qu trnh ct ni, s sp xp li lm snARN U6 bt cp vi snARN
U2 v snARN U5 tng tc vi 4 nucleotid ca exon, thay th snARN U1 t pre - mARN. snARN U2
cn li to cp base vi intron gn vng giu pyrimidin.
Lc ny, phn ng chuyn v ester th nht xy ra do phc hp U2 - U6 xc tc.
- snRNP U5 tng tc vi phn intron ca v tr nhn v U2, xc tc phn ng ct lin kt intron -

exon v ni 2 exon. Phn ng chuyn v ester th 2 xy ra.
- Spliceosom c tho ri, gii phng U1, U2, U4, U5, U6.
- Si i intron c tho xon v thoi ho.
Intron nhm II: t gp
y l phn ng t ct ni, intron ng vai tr nh mt enzym - gi l ribozym.
Ct ni cho
Cc phn ng ct ni trn xy ra trn cng mt phn t ARN, gi l ct ni kiu cis; nu xy ra gia

cc phn t gi l ct ni kiu trans.
u tin, v tr nhnh A si ln ti vng giu pyrimidin u 5 ca exon m ho lin kt vi vng

3 ca ARN dn, gii phng "tiu exon". Sau , u 3 ca tiu exon t do lin kt vi u 5 ca exon
m ho, ct b on nhnh cha vng 3 ca ARN dn v vng 5 m ho ca bn sao nguyn thu. Cc
bc ny tng ng vi phn ng chuyn v ester ho I, II trong ct ni pre - mARN t bo nhn tht
bc cao.
Hnh 3.11. Ct ni cho ca ARN dn vi exon m ho protein bn sao nguyn thu

polycistron loi Trypanosoma nh hai phn ng chuyn v ester ho
3.2.5.4. X l tARN
Pre - tARN c ct v loi b intron tr thnh tARN trng thnh.
X l u tn cng 5' v 3'
- X l u 5: enzym xc tc l ribonuclease (RNseP)
- X l u 3: kt hp endonuclease v exonuclease. Nu u tn cng 3 cha c trnh t CCA th

tARN nucleotidyl transferase s b sung trnh t ny.
Ct ni tARN nm men
- ARN endonuclease (RNse) ct tin tARN ti 2 u ca intron, to nhm 5 - OH v 23P vng .
- 2 na phn t tARN cng tn ti nh lin kt hydro.
- Pi t GTP gn vo nhm 5 - OH ca u 3 exon nh enzym GTP kinase.
- 2 na phn t tARN sau lin kt vi nhau nh enzym ARN ligase.
- Vng 23P c m nh enzym cyclic phosphodiesterase (l mt phn ca phn ng kt ni), gii

phng 2Pi.
- Phosphatase loi b 2Pi.
Hnh 3.12. Ct ni tARN nm men
3.2.5.5. X l rARN
Ct ni theo c ch nhm intron I t bo nhn tht bc thp, nm v 3 gen ca thc khun T4

E.coli. C 2 tnh cht chung:
- T ct ni
- Cu trc bc 2 ca vng 9
Ct ni loi tetrahymena
Tin rARN 35S c x l thnh 18S, 5,8S, 26SrARN. mt vi chng Tetrahymena, 26S c mt
intron gm 400 base. Intron ny c th t ct ni nhng i hi s c mt ca cc ion v nucleotid G
(GTP, GDP, GMP)
Phn ng t ct ni: gm 3 bc
1. G 3 - OH gn u 5 ca intron.
2. 3 - OH ca exon A gn u 5 ca exon B: cc exon c ni vi nhau.
3. Intron mch thng vi u 5 tn cng bng G (414) s c ng vng.
ng vng intron mch thng
4. Nhm 3 - OH ca intron ti G (414) gn vi pA (16) hay pU (20), to vng intron v khi u 5

15 hay 19 nucleotid.
5. Nc thu phn vng intron ti cng v tr.
6. Nu qu trnh ng vng u tin xy ra ti pA (16) th nhm 3 - OH ca intron gn vi pU (20),

to vng intron v ct 3 nucleotid mch thng.
7. Nc thu phn vng intron ti cng v tr, to L - 19ARN. L - 19ARN c hot tnh enzym xc
tc phn ng ct v ni cc oligonucleotid.
Vai tr cu trc bc 2 ca intron ARN, vng 9
8. Nhnh P1 (exon 1 v IGS InteARNl Guide Sequence) ng vai tr quan trng trong bc 1 ca
phn ng ct ni.
9. Nhnh P4 v P7 ng vai tr quan trng nht trong bc 2 ca phn ng ct ni, ngoi ra cn c

P3, P6 v 2 base u 3 ca intron.
Vai tr xc tc ca L - 19ARN
10. Gn ARN vi IGS v tr gn kt.
11. Gn G (414) hay G t do vo v tr gn kt.
Tm tt
ARN gm mt chui n ca cc ribonucleotid ni vi nhau bi cc lin kt phosphodiester. Trnh

t nucleotid ca ARN b sung vi trnh t mt mch n (mch khun) ca ADN xon kp.
C 3 loi ARN chnh: mARN, rARN v tARN. Tt c u tham gia vo qu trnh dch m, nhng
ch c mARN mang thng tin m ho cho cu trc c bn ca phn t protein. rARN c vai tr trong
vic cung cp ni tng hp protein, ribosom. tARN lm nhim v vn chuyn acid amin n ribosom,
tng tc vi aminoacyl - tARN synthetase c hiu v kt hp vi ribosom.
Tt c ARN u c to ra t s sa i v bin i ca nhng tin ARN ln. Hn na, t bo

nhn tht cn c mt s phn t ARN nh vi nhng chc nng khc nhau bao gm thnh phn ca
enzym ribonuclease, di chuyn intron, kt ni exon v mt yu t kim sot vic tng hp ca protein
bi tit.
CU HI
1. Tnh cht no khng phi ca tt c ARN?
a) Mch n polynucleotid
b) ng pentose (5C) l ribose
c) Ngoi A, G, C th uracil thay cho thymin
d) c tng hp t trong nhn
e) C lin kt hydro gia A = T
2. Cu to t 34 phn t protein, 1 phn t rARN 23S, 1 phn t rARN 5S l tiu n v:
a) 50S b) 30S
c) 60S d) 40S
e) 70S
3. Cu to t 45 phn t protein, 1 rARN 28S, 1 phn t rARN 5.8S, 1 phn t rARN 5S l tiu n
v:
a) 60S b) 40S
c) 50S d) 30S
e) 70S
4. Tiu n v 40S ca t bo nhn tht cu to t:
a) 34 phn t protein + 1 rARN 23S, 1 rARN 5S
b) 21 phn t protein + 1 rARN 16S
c) 45 phn t protein + 1 rARN 28S, rARN 5,8S, rARN 5S
d) 33 phn t protein + 1 rARN 18S
e) 45 phn t protein + 1 rARN 23S + 1 rARN 5S
5. Tiu n v 30S ca t bo nhn nguyn thu cu to t:
a) 34 phn t protein + 1 rARN 23S, 1 rARN 5S
b) 21 phn t protein + 1 rARN 16S
c) 45 phn t protein + 1 rARN 28S, rARN 5.8S, rARN 5S
d) 33 phn t protein + 1 rARN 18S
e) 45 phn t protein + 1 rARN 23S + 1 rARN 5S
6. Qu trnh methyl ho nh ARN - methylase ch xy ra :
a) mARN b) Pre - rARN
c) tARN d) scARN
e) snARN
7. Tnh cht no khng c hiu cho tARN?
a) Chiu di khong 73 93 nucleotid
b) Mch n cun hnh l ch ba
c) u mt 3 kt thc CCA gn acid amin
d) u mt 5 kt thc G
e) Mt loi tARN c th mang nhiu loi acid amin khc nhau
8. Phn ng no khng phi ca tARN trong qu trnh sinh tng hp protein?
a) Aminoacyl ho
b) Formyl ho tARN m u
c) Gn nhng yu t kt thc
d) Gn ribosom
e) Nhn din codon anticodon
9. Loi snRNP no tham gia vo vic sa i hnARN thnh mARN hon chnh?
a) U1, U3 b) U4, U5
c) U6, U7 d) U1, U2
e) U1, U4
10. t bo nhn tht mARN sau khi c phin m phi tri qua:
a) Gn chp b) Gn ui polyA
c) Ct ni loi intron d) a, b
e) a, b v c.
Bi 4
S PHIN M V M DI TRUYN
MC TIU
- Trnh by c qu trnh phin m t bo nhn nguyn thu v t bo nhn tht.
- M t c s phin m c iu ho.
- Trnh by c s phin m ngc, nu c ngha ca n trong chu trnh sng ca

Retrovirus.
- Nu c bn cht ca m di truyn v cch nhn bit.
- Phn bit c cc loi ARN polymerase v cu trc v chc nng.
4.1. M U
ADN trong nhim sc th ng vai tr lu tr thng tin di truyn ca t bo. N quy nh trnh t
acid amin ca hng ngn protein khc nhau. Tnh ng nht v s lng cc protein ny quyt nh tnh
cht sinh ho cn bn ca t bo. ADN khng trc tip sp xp cc acid amin cho s polymer ho thnh
protein m l mARN, l bn sao b sung vi mt mch polynucleotid ca gen. Qu trnh to ra cc ARN
c gi l s phin m. Tn hiu c m ho trong ADN hng dn enzym thc hin phin m, ni
m u, ni kt thc.
S iu ho mc phin m l s kim sot quan trng hng u trong biu hin gen. ARN c
phin m t gen ph thuc vo mt vng nht nh c gi l promoter (vng khi ng). Bn cnh
vng khi ng, cn c mt vi trnh t khc trn ADN cng c vai tr quan trng trong vic iu ho
biu hin gen. Vn ny c nghin cu k v biu hin r t bo t bo nhn nguyn thu, cn

t bo nhn tht ch mi c nghin cu t khi c k thut ADN ti t hp nhng cng thy c
mt s khc bit vi t bo nhn nguyn thu.
S nhn din cc acid amin lin kt chng vi nhau thnh chui polypeptid c xc nh bi cc
trnh t ribonucleotid trong mARN. l mARN c dch m t "ngn ng" nucleotid ca ADN. M
di truyn m t mi quan h gia 2 ngn ng ny.
4.2. NGUYN TC CHUNG
Qu trnh truyn thng tin di truyn t ADN sang ARN c gi l s phin m. Tham gia vo qu
trnh phin m c h thng enzym ARN polymerase, ng ra, tn chnh xc l ARN polymerase ph
thuc ADN. ADN th hin kh nng d xc tc tc l lm khun tng hp nn mt phn t khc.
S phin m thc hin theo cc nguyn tc:
- Ch 1 trong 2 mch ca phn t ADN dng lm khun tng hp ARN (hnh 4.1)
- ARN polymerase bm vo ADN lm tch mch v di chuyn theo hng 3 5 trn ADN cho
"ARN" c tng hp theo hng 5 3.
- ARN polymerase gn vo ADN gy ra hin tng "chy" ti ch bn trong chui xon kp ADN.
Kt qu lm t cc lin kt hydro gia cc i base b sung. C cht cho ARN polymerase l cc
ribonucleotid 5 - triphosphat ATP, GTP, CTP v UTP. Mt ion kim loi ho tr 2 hoc l Mg2+ hoc l
Mn2+ ng vai tr co - factor.
Cu trc ca phn t ARN thng bt u bng 1 nucleotid purin (ATP hay GTP) b sung cho
nucleotid im bt u phin m ca mch khun mu ADN. NTP th 2 c a vo khun mu
ADN (li theo nguyn tc b sung cp base) v lin kt 3 - 5 phosphodiester c hnh thnh gia
nhm 3 - OH ca NTP u tin v nhm phosphat ( ) ca NTP th 2.
Hnh 4.1. Qu trnh chuyn thng tin t ADN n protein
Phn ng xy ra theo pha thu phn pyrophosphat va c gii phng. Bng cch ny ARN c
tip tc ko di v mt t hp lai ADN - ARN c hnh thnh. Chui ARN ko di theo hng 5 - 3
v ARN polymerase cng vi vng si kp ADN dui xon (cn gi l bong bng phin m) chuyn
ng dc theo si khun mu theo hng 3 5. Khi t hp lai ADN - ARN t c chiu di 12
base, u tn 5 ca chui ARN mi hnh thnh c tch ra t mch ADN b sung vi n v 2 mch
kp ca ADN ban u phc hi li. ARN polymerase tip tc di chuyn dc theo mch ADN khun
mu n khi gp im kt thc, l tn hiu c m ho trong mt trnh t ADN quy nh chm dt
s ni di ca mch. ARN polymerase v chui ARN hon chnh c tch ra khi khun mu ADN.
ARN polymerase thc hin s lin kt cho 4 loi nucleotid triphosphat ging nhau, khng phn bit
loi ny, loi kia. Khc vi s sao chp ADN, mi khng cn thit cho s phin m.
Trong a s trng hp, ch c mt trong 2 mch ca ADN c phin m v s phin m ny c

gi l phin m bt i xng. Cng c nhng trng hp trong s phin m xy ra c 2 mch
ADN, l s phin m i xng, v d, s phin m ADN ty th t bo th, trong c 2 mch ca
b gen vng 16 kb u c phin m ra ARN. Tuy nhin, sau 1 trong 2 bn sao ny b thoi ho.
Khc vi ADN polymerase, ARN polymerase khng c mt hot tnh sa sai no, v nhng
nucleotid no kt hp khng chnh xc thng c thay th ngay bng nhng nucleotid ph hp. Hn
na, nu c li him hoi no xy ra th cng khng di truyn c v ARN khng phi l ni lu tr
thng tin di truyn.
Chui ADN c phin m to ARN c gi l 1 n v phin m v sn phm l bn sao nguyn

nguyn thu. Bn sao nguyn thu ny c bin i trc khi m nhn vai tr chc nng ca n. Tt
c cc loi ARN t bo (mARN, tARN, rARN v cc ARN nh) u c phin m t ADN.
4.3. S PHIN M T BO NHN NGUYN THU
S phin m t bo nhn nguyn thu c nhng c im sau:
- Ch mt loi ARN polymerase chu trch nhim tng hp tt c cc loi ARN.
- mARN thng cha thng tin nhiu gen ni tip nhau (polycistron mARN).
- Qu trnh tng hp mARN c tin hnh khi ARN polymerase bm vo vng khi ng
(promoter). Khi gn vo, polymerase khng tng hp mARN ngay. S tng hp ch bt u t du xut
pht, thng l TAC nm cch 7 - 8 base pha sau ch bm v pha u 5 ca mch khun.
- phn ln t bo nhn nguyn thu, qu trnh tng hp tip tc n khi c qua du kt thc. Kt
thc phin m, ARN polymerase v mARN tch ri khi ADN.
- Qu trnh phin m v dch m xy ra ng thi vi nhau.
Hu ht s hiu bit v qu trnh phin m t bo nhn nguyn thu xut pht t s nghin cu
trn E. coli v thc khun ca n. C ch ca s phin m E. coli tng t hu ht t bo nhn
nguyn thu, ngoi tr Archaebacteria (c intron trong ADN).
Nhim sc th n bi ca E. coli l mt vng khp kn cha 4.106 bp (cp base) cng vi protein v
ARN. ADN ny c cu hnh tho xon cc i (negatievely supercoiled conformation). In vivo, nhim
sc th l mt cu trc lng lo gi l nucleoide v c gn vo mng t bo cht. B gen ca E. coli
cha khong 3000 gen cu trc khc nhau, hu ht c mt trong mt bn sao duy nht, sp xp thnh
100 n v phin m.
4.3.1. ARN polymerase ca E. coli
L mt protein phc hp cha ti 5 tiu n v gm 4 loi v c trng lng phn t khong

450000. Ton b enzym c gi l holoenzym cha cc tiu n v ' .
Tiu n v (sigma) c th tch ra thun nghch t holoenzym v c phn tch bng sc k trn
ct phosphocellulose:
4.3.2. Promoter
Phn ln promoter E. coli v c bn u ging nhau. Nu k hiu base u tin phin m thnh
mARN (thng l Adenin) l + 1, th cc base theo hng ngc li vi chiu phin m k hiu " - ".
Promoter gm hai trnh t. l trnh t TATAAT, tn cng lun lun l base T, cch im xut
pht 6 - 8 base v c base trung tm l 10. Trnh t ny cn c gi l hp TATA hay hp Pribnow;
trnh t th 2 TTGACA c base trung tm - 35 (hnh 4.2). Hai trnh t ny chu trch nhim cho php
ARN polymerase gn vo vng khi ng phin m. Vng 35 tham gia vo vic gn khi u ARN
polymerase (bng 4.1). Khng phi promoter no cng c cu to ging ht cc trnh t ny, m c mt
t sai lch nh, do ngi ta gi trnh t ny l trnh t chung (consensus).
Bng 4.1. Cc trnh t chung -10 v -35 ca mt s promoter ca E. coli
Promoter - 35 - 10 im xut pht
Consensus TTGACA TATAAT
lac P TTTACA 18 bp TATGTT 6 bp A
Gal P (c ch ng ho) TATGGT 6A

1
Gal P (c ch d ho) GTCAC 16 TATGCT 6A

2
AraC CTGACA 17 TGTCAT 6G
AraBAD CTGACG 18 TACTGT 8A
Trp TTGACA 17 TTAACT 7A
Hnh 4.2. Cu hnh vng - 10 v - 35 trn ADN
Chc nng quan trng ca hp -10 v hp -35 c pht hin t hiu ng t bin (mt base c
thay th bi mt base khc) trong qu trnh phin m t promoter (hnh 4.3). Nhiu t bin nh vy c
ch tc khc s phin m ca gen. Tht ra, vi vng khi ng (promoters) E. coli c trnh t chung
rt ging vi trnh t chung l tng. Do , phn bit c chng c th da vo cng
(strength) hoc tn sut khi ng phin m. V d, t bo E. coli cha khong 12 phn t ca lac
repressor (th c ch lac - operon) v gen ca n c phin m ch mt ln mi 30 pht. Mt khc,
ARN ribosom, do i hi s lng ln, v khong 10000 ribosom phi c hnh thnh sau mi th h
t bo (khong 20 pht). Nn mi gen - rARN c phin m khong 2 giy mt ln. S khc bit ny
do cng vng khi ng.
Hnh 4.3. Cc v tr t bin im tc ng ln chc nng

vng khi ng E. coli
4.3.3. Cc promoter shock nhit
Mi cho ti gn y, ngi ta vn cho rng ARN polymerase E. coli c mt loi holoenzym duy
nht c kh nng phin m tt c cc gen khc nhau. Tuy nhin, nhng nghin cu gn y trn s p
ng shock nhit cho thy rng kh nng phin m cc gen khc nhau c th t c do sinh ra cc
tiu n v khc nhau. Ngi ta chng minh s p ng shock nhit c lin quan n vic tng hp
nhng protein mi khi c s gin on nhit mi trng nui cy vi khun hay t bo. S p ng ny
bin thin rt rng t bo nhn tht v t bo nhn nguyn thu. Nhng protein mi ny, c gi l
protein shock nhit, c xem l bo v t bo chng li s bin i nhit.
E. coli c khong 17 protein shock nhit m s biu hin ca chng c to ra bi sn phm ca

gen hptR. Gen ny p ng vi shock nhit v m ho cho chui polypeptid c Mr 32 000, ng vai tr
nh mt yu t bin i. Yu t ny (c gi 32) kt hp vi phn li bnh thng ca ARN
polymerase to nn holoenzym 232. Enzym ny ch bm vo nhng promoter ca gen shock
nhit, ch khng bm vo promoter ca cc gen bnh thng v c trnh t vng - 35 di hn vi mt s
khc bit nh.
Hnh 4.4. Cc trnh t chung ca promoter sc nhit E. coli
Vi khun Bacillus subtilis cng s dng yu t bin t dng sinh dng sang dng bo t.
Nhiu ADN thc khun th m ho ARN polymerase c trng cho ring chng. Mt s promoter
nh vy c s dng trong vic to dng plasmid nhm sn xut mt s lng ln mARN t mt gen
quan tm. Nhng bn m sao nh vy c th c s dng nh nhng on mi hoc c th c tng
hp in vitro thnh mt sn phm protein cn thit.
Trnh t chung ca promoter sc nhit c sp xp cho 6 gen sc nhit khc nhau. Nhng ch ci
dng trn ch s tng ng mnh, hng di tng ng yu, n l nucleotide bt k. V tr bt u
phin m k hiu +1.
Ngi ta thng dng h thng promoter v ARN polymerase ly t thc khun th SP6. SP6 l mt
thc khun th ca Salmonella typhimurium, m ho c mt ARN - polymerase virus c hiu. Cc
promoter ca t bo ch v cc promoter c trong plasmid ly trch t E. coli u khng c SP6
polymerase nhn bit. Promoter SP6 c chn vo vector plasmid pha trc polylinker (polylinker l
on ADN nhn to cha nhiu v tr nhn bit duy nht ca cc enzym ct hn ch). on ADN cn
sao chp c chn vo polylinker v plasmid ti t hp c bin np vo E. coli. Plasmid cha ADN
l c "lm dui thng" bi mt enzym ct hn ch. Plasmid dui thng ny c vi SP6
polymerase v cc NTP. Cc plasmid - ADN c loi khi cc bn sao mARN bng cch vi DNase
(hnh 4.5).
Hnh 4.5. Dng SP6 promoter v ARN polymerase

ca n nhn bn sao m cn thit
4.3.4. Khi u v ni di chui ARN
Vic gn ca holoenzym ARN polymerase vo promoter to ra phc hp khi ng ng c cu

hnh ADN xon kp (hnh 4.6). Cc im tip xc ch yu gia promoter v ARN polymerase nm
cc vng -10 v -35. Phc hp ng nhanh chng bin thnh phc hp m do cc lin kt hydro b
enzym ct trn mt vng bao gm 17 bp. Si ADN c tch ra nh s tho xon 1,6 vng ca xon
kp B - ADN. Nucleotid triphosphat u tin (thng l ATP hoc GTP) gn vo tiu n v ca
polymerase, hng ng vo base b sung v tr +1 ca si khun mu. Mt nucleotid triphosphat th
2 tip theo c kt ni v hnh thnh lin kt phosphodiester u tin. Nh vy, chui mARN pht
trin mang triphosphat u 5 v theo hng 5 3. V im ny, phin m ging vi sao chp
ADN.
Hnh 4.6. S khi u v ni di chui ARN bi ARN polymerase ca E. coli
(a) Holoenzym gn vo promoter to thnh phc hp khi ng ng. Cc trnh t - 10 v - 35 ng vai tr ch

yu cho s gn ny. (b) ARN polymerase lm chy ti ch chui xon kp hnh thnh nn phc hp khi ng
m. Triphosphat nucleotide u tin c kt ni v hnh thnh cp base ti im xut pht + 1. S ko di
chui ARN bt u khi cc NTP polymer ho vo u 3 - OH ca ng ribose ca chui. Bong bng phin m
di chuyn dc theo si khun ADN theo hng 3 5. Khi kt dnh c khong 12 nucleotide vo chui th
tiu n v tch ra li phn li ca enzym hon tt qu trnh ni di. Tiu n v li c th lin kt vi phn
li t do trong dch bo tng tham gia vo vng khi u mi.
E. coli, hu ht mARN c i sng bn hu c tnh bng pht. Chng nhanh chng b thoi ho
bi h thng enzym endonuclease v exonuclease. Tht ra, phn ln mARN c trong t bo cht bt
k thi im no cng u l nhng chui mi sinh. Chng gn vo phc hp ADN khun - ARN
polymerase. Ribosom gn vo v tr gn ribosom gn u 5 ca mARN va mi ri phc hp ADN
khun - ARN polymerase. Do , s phin m v s dch m t bo nhn nguyn thu c thc hin
gn nh song song vi nhau. Chnh iu ny lm xut hin mt dng kim sot phin m c gi l
phin m suy gim.
4.3.5. Kt thc qu trnh tng hp ARN
Chui ARN ni di cho n khi gp phi mt trnh t chuyn bit trong chui ADN. Chui ny
dng kt thc qu trnh tng hp ARN. E. coli c 2 c ch kt thc: mt c ch ph thuc vo protein
ph km c gi l (yu t rho) v mt c ch khc khng ph thuc vo cc yu t ny.
4.3.5.1. Kt thc c lp vi yu t rho
Qu trnh ny c trng bi s hin din ca mt trnh t giu GC trn ADN. Trnh t ny i xng
2 bn (dyad symmetry), c thm 5 hoc 6 adenin km theo. ARN c phin m t trnh t ny c th
hnh thnh mt nt vng hay mt cu trc kp tc do cc lin kt hydro ni phn t gia cc base b
sung (hnh 4.7). C th s hnh thnh xon kp ARN trong cu trc nt ny, thch hp cho vic hnh
thnh ADN - ARN lai gia si ADN phin m v chui ARN mi sinh. Nh vy, phn dn 5 ca nt
ko theo na ui 3 t ADN, ch li trnh t oligo (U) tip tc ni di theo ADN. Cc dng lai
oligoA - oligoU cho thy s bt cp t bn vng nht v ARN c th tch khi ADN. Sau , si kp
ADN c hnh thnh trong "bong bng phin m" v phn li ca enzym polymerase, c i lc thp
i vi ADN kp, c phng thch ra.
Hnh 4.7. Cu trc bc 2 c gi thit ca u 3 ca bn sao ARN

t trp operon ca E. coli
Nt vng hay kp tc v trnh t UUUU to nn im kt thc
4.3.5.2. Kt thc l thuc yu t rho
S kt thc ny cng cn s c mt ca nt v vng ngay trc u ko di ca ARN. Nhng khng

c trnh t oligo (U). Yu t l mt protein gm 6 tiu n v ging nhau c Mr 46 000 v c i lc
cao vi chui ARN n.
Khi bm dnh vo ARN, yu t thu gii ATP v nng lng t do c gii phng, gip n di
chuyn dc theo chui ARN mi sinh, hng ti bong bng phin m (hnh 4.8). Sau , yu t tch
i ADN - ARN lai bng mt c ch cha r, phng thch mARN vo t bo cht.
Hnh 4.8. M hnh gi thit cho s kt thc phin m E. coli bi yu t rho
4.4. S PHIN M T BO NHN THT
Phin m t bo nhn tht c cc c im sau:
- ARN polymerase II chu trch nhim tng hp mARN, ARN polymerase I tng hp rARN, ARN
polymerase III phin m cho ra cc ARN c kch thc nh, tARN, ARN 5S
- mARN cha thng tin ca mt gen (monocistron mARN)
- Qu trnh phin m phc tp hn. u 5 mARN c gn thm "chp" (CAP) l 7 -

methylguanosine, cn cui mARN pha 3 c thm "ui" polyadenin di 100 - 200 nucleotid
- Bn phin m u tin cn gi l tin mARN khng c s dng trc tip m phi qua bin i.
4.4.1. Cc gen gin on
T nm 1977, ngi ta pht hin nhiu gen ca t bo nhn tht c tnh gin on. Trn gen, cc
on m ho cho protein c gi l exon xen k vi cc on khng m ho c gi l intron. Bn
phin m u tin tc l tin mARN cha c trnh t phin m ca exon v intron. Tip theo cc intron
tc cc on khng m ho protein c ct ri ra, cn cc exon m ho protein c ni lin li vi
nhau to ARN trng thnh. Qu trnh ny c gi l ct ni (hnh 4.9).
Khi mch mARN ang c ni di khong 20 - 30 nucleotid th u 5P, enzym s ni thm vo

gc 7 - methylguanylate. Chp ny gn vo u 5P to nn lin kt 5P - 5P.
Mt on ngn ca ui 3 pre - mARN, ti phn khng m ho sau b kt thc, b ct ni ui

poly A.
Phc hp ribonucleoprotein (snRNP) ca nhn t bo thc hin qu trnh ct ni (splicing) to cu

trc khng gian thun tin cho cc u exon gn nhau v xc tc phn ng kt ni.
Sau splicing, mARN va mi trng thnh khng cn intron, qua l mng nhn ra t bo cht
dch m.
Khng phi mARN ca t bo nhn tht no cng cn gn chp, ni ui v ct ni.
Ch c mt t l nh (khong 10%) b gen ca t bo nhn tht l phin m c. Phn ln cn li

ca b gen trng thi si nhim sc ng xon mnh (d nhim sc) nn tr vi s phin m. Phin m
ch xy ra cc nguyn nhim sc xon lng lo.
Hnh 4.9. Phin m gen gin on t bo nhn tht
4.4.2. ARN polymerase ca t bo nhn tht
Nhn ca t bo nhn tht c 3 loi ARN polymerase (I, II, III) (bng 4.2).
Bng 4.2. Cc ARN polymerase nhn ca t bo nhn tht
Loi V tr Bn sao Hiu ng - amanitin
Polymerase I Nhn con Tin rARN ln Khng nhy cm
Polymerase II Dch nhn Tin mARN (hn ARN) Rt nhy cm
Polymerase III Dch nhn 5S rARN, tARN v cc Nhy cm trung bnh

ARN nh khc
Polymerase I trong nhn, phin m cc gen ARN ribosom. Bn sao ch yu l pre - rARN.
Polymerase II tm thy trong dch nhn, rt nhy cm vi s c ch ca - amanitin (khong 50% c
ch nng 10-8 M). N phin m cc gen m ho protein v cc bn sao s cp khng ng nht
(hnARN), l tin cht ca mARN t bo cht. Polymerase III cng trong dch nhn, phin m cc gen
5S rARN v gen tARN. N nhy cm trung bnh vi s c ch ca - amanitin (khong 50% c ch
10-6 M). Phn ng c xc tc bi cc polymerase t bo nhn tht, v mt ho sinh cng ging vi
cc phn ng c xc tc bi cc enzym E.coli.
C 3 polymerase ca t bo nhn tht u c phn t lng ln vi M vt qu 500000. Chng c 2

tiu n v ln hn 100000, mi phn l mt polymerase chuyn bit v c ti 12 tiu n v nh hn,
trong c ti mt vi phn khng gp c 3 polymerase. Chc nng chnh xc ca cc tiu n v
khc nhau cha c bit ht. Khc vi cc polymerase vi khun, polymerase nhn t bo nhn tht
khng c kh nng khi u phin m cc vng tng ng vi cc v tr xut pht in vivo, khi enzym
tinh khit vi ADN in vitro.
4.4.3. S phin m do ARN polymerase I
Cc gen ARN ribosom thng l cc gen c sao chp mnh nht trong t bo, phn nh nhu cu
cao v rARN. Si nhim sc (chromatin) cha cc gen ribosom, hon ton khng to nucleosom. Cc
trnh t sao chp cc gen rARN c bo lu mc cao trong cc chng sinh vt, chnh v th tht
khng l khi tm thy c cc trnh t chuyn bit cho tng loi pha trc cc im xut pht phin m.
l mt vng khng phin m nm gia cc trnh t c phin m ca cc gen rARN ca Xenopus
laevis (cc chu Phi) v cc trnh t cn thit cho s phin m bi polymerase I. Cc trnh t cn thit
ny c xc nh bng tc ng xo b (hnh 4.10).
Promoter thc gm khong 150 bp c tm thy gia cc v tr - 146 v + 6. Tuy nhin, cc bn sao
chp hon chnh t di ca promoter ny c lp li cc v tr - 1200 v - 2300 bp trong vng v
tinh (vng khng phin m) v ci rt nhiu bn sao 60 hoc 81 bp ca promoter thc. C l iu ny
lin quan ti mt yu t gii hn s lng. Cc bn sao lp li 60/81 bp kch thch rt nhiu s phin m
ca promoter thc. Cc promoter v tinh ny c kh nng khi ng qu trnh tng hp cc bn m sao
rARN. Cc phn t polymerase "xp hng" trong vng v tinh trc khi chuyn ng ti promoter thc.
Hnh 4.10. Cc phn t promoter trong vng v tinh khng phin m ca

cc gen rARN Xenopus laevis
vung en: cc trnh t lp li 60 hoc 80 bp c trong promoter thc. Cc v tr - 1200 v -

2300 l cc bn sao hon chnh ca cc promoter thc v tr + 6 n - 142.
ngi, cc rARN c phin m bi ARN polymerase I. Bn phin m u tin l tin rARN 45S
(pre - rARN), sau ct ni to ra 3 loi 28S, 18S v 5,8 S. Cc gen ny khng phn tn m xp thnh
cm, mi cm c th hn 200 bn sao. ngi, cc cm ny nm trn vai ngn ca cc nhim sc th
tm u 13, 14, 15, 21 v 22. Cc nhm ny xp quanh yu t t chc hch nhn hnh thnh eo th cp
v gian k to nn hch nhn.
4.4.4. S phin m bi polymerase II
Cc gen c phin m bi polymerase II to ra mARN a dng hn nhiu so vi cc gen rARN

ng nht.
Tr cc gen m ho cho protein histon, th cc gen c phin m bi ARN polymerase II u c

trnh t duy nht hoc vi s t bn sao. Cc gen ny hu nh ch m ho cho mt loi protein.
Mt s gen c bn sao th 2 trong qu trnh tin ho. C 2 bn sao c th chuyn i b tr nhau,

nh trng hp cc gen - globine. Cc gen m ho protein ny biu hin thng xuyn cc m (gen
gi nh) v c th ch biu hin mt m no y ca c th a bo (a phn l gen iu ho) hoc iu
ho bi s c hay thiu c cht ring bit, nh hormon hoc cht kch thch mi sinh. Cc gen lin quan
n s chuyn ho galactose trong nm men l mt th d.
Cc promoter ca ARN polymerase II nm pha 5 ca im bt u phin m. Cc trnh t tip

theo rt c ngha i vi chc nng ca promoter (hnh 4.11).
Hnh 4.11. Cc nt chung ca promoter ca ARN polymerase II t bo nhn tht bc cao,

s lng v v tr ca cc hp GC rt khc nhau
Hp TATA hay hp Goldberg - Hogness c nt chung, nhng khng c ngha l ph bin v c mt

trnh t chung TATAAT. N nm cch khong 25 bp ngc chiu vi v tr bt u sao m cc t bo
nhn tht bc cao. Nhiu hp TATA gn km vi cc trnh t giu GC. Cc trnh t ny ng vai tr rt
quan trng trong vic la chn v tr bt u phin m.
Hp CAAT (nm cch v tr phin m 80 bp), c trnh t chung l GGCAATCT.
Hp GC hay thay i v tr, c trnh t chung l GGGCCGGG.
Ngi ta cho rng cc trnh t chung ny c chc nng lin kt cc yu t phin m chuyn bit hn
l gn trc tip vi ARN polymerase II. V d, cc t bo ca ng vt c v cha 1 protein gi l SP1
c Mr 100 000, cn cho s phin m ca cc gen c hp GC trong promoter. Cc trnh t khi ng
khc nhau nm rt gn vi im bt u phin m l cn thit, nhng thng vn khng biu hin
cc gen ARN polymerase phin m. C cc phn t c a thm vo, gi l cc trnh t tng cng.
Chng khng c kh nng khi ng chnh mnh, nhng li c kh nng kch thch phin m mnh v c
th tc ng cc khong cch ng k (n vi kb ADN). Trnh t tng cng c hiu qu nh nhau
i vi cc gen cu trc do chng iu ho v c th nm rt gn u 3, 5 hoc nm ngay trong mt
cistron ca gen cu trc.
Trnh t tng cng lun th hin tnh chuyn bit, do khng lin quan ti vic iu ho biu
hin gen trong qu trnh pht trin ca c th. V d, trnh t tng cng min dch globulin ch hot
ng trong - lymphocyte.
Mt trong nhng v d v hot ng tng cng c nghin cu trong iu ho phin m gen p
ng vi hormon steroid. Gn steroid vo th th protein tan trong nc, cho phc hp ny li gn

vo cc trnh t tng cng lm gia tng hot ng cc gen p ng steroid (hnh 4.12).
Hnh 4.12. Hot ho qu trnh phin m ca cc gen

p ng hormon steroid (glucocorticoid)
Cc bn m sao ca ARN polymerase II u l nhng ARN nhn khng ng nht (hnARN). Chng
c phn t lng rt ln (n 10 kb), v cha cc intron trong ADN. a phn cc bn m sao u cha
trnh t AAUAAA gm 20 nucleotid t u 3. Trnh t ny rt quan trng v: n c nhn bit bi
mt endonuclease c hiu ct ARN mi sinh khi polymerase II v n l du hiu c nhn bit
bi mt enzym poly(A) polymerase nhm gn ui 3 ARN.
4.4.5. S phin m bi ARN polymerase III
ARN polymerase III chu trch nhim phin m ra rARN 5S, tARN v mt s ARN nh ca nhn v
ca t bo cht. Cc gen do ARN polymerase III phin m thng cha di 300 nucleotid. Gen 5S
rARN khng cha intron, trong khi , nhiu gen tARN cha cc intron nh.
Tham gia iu ho tng hp cc rARN 5S ca non bo X. laevis, ngoi ARN polymerase III, cn c
cc yu t phin m khc l TFIIIA, TFIIIB v TFIIIC. Hai yu t TFIIIB v C cng cn cho s phin
m cc gen khc bi ARN polymerase III (chng hn cc gen tARN), trong khi TFIIIA (mt protein
c Mr 40000) th chuyn bit cho non bo v gn vi vng ni kim sot ca gen rARN 5S. Sau ,
hng ARN polymerase III tng hp khong 50 nucleotid (hnh 4.13).
Hnh 4.13. Kim sot qu trnh tng hp rARN 5S non bo X. laevis
TFIIIA thng gn bm vo rARN 5S t do trong t bo cht non bo hnh thnh nn phc hp 7S

ribonucleoprotein (7S RNP). Nu vic tng hp cc thnh phn khc ca ribosomnh cc ARN 28S, 18
S v protein ribosom din ra sau qu trnh tng hp rARN 5S, th gn nh tt c TFIIIA gn trong
phc hp 7S RNP v do khng cn TFIIIA cho s phin m. Chnh v vy, s tng hp rARN 5S b
gim xung cho n khi TFIIIA c gii phng tr li. Do , c th xem s tng hp rARN 5S non
bo hon ton l qu trnh iu ho.
4.5. PHIN M NGC RETROVIRUS
Retrovirus l cc virus m vt liu di truyn ca chng l ARN, v d virus lm suy gim min dch
(HIV) gy ra hi chng suy gim min dch mc phi ngi (AIDS). S tng hp ADN t khun mu
ARN c xc tc bi cc enzym phin m ngc. Enzym ny bt u qu trnh phin m bng cc
dNTP nh l cht nn (hnh 4.14). Khc vi ARN polymerase, enzym phin m ngc phi dng ti
mi. on mi l mt tARN ca t bo ch, gn kt vo u 3ca retrovirus. Chui kp u tin l
chui lai ARN - ADN. ARN virus trong chui lai b hu bi RNaseH. Sau , mt mch ADN va c
tng hp to ra chui kp ADN. Chui kp li tch hp vo ADN k ch.
Enzym phin m ngc ca HIV l mt protein gm 2 tiu n v. Mi tiu n v cha 560 acid
amin. Enzym phin m ngc c mt vai tr quan trng trong vic tng hp ADN b sung (cADN)
trong qu trnh iu khin gen. S tch hp ca virus vo b gen t bo ch thng c n nh (cc
provirus khng bao gi b ct ri nh prophage). Hin nay, nhiu ngi cho rng cc virus c th tc
ng n s biu hin ca cc gen ung th.
Hnh 4.14. Sao chp ngc si n ARN ca Retrovirus thnh si kp ADN
4.6. M DI TRUYN
Ta bit, cc acid amin trong phn t protein c m ho bng nhm cc nucleotid trn phn t
ADN. C tt c 4 loi base, nu cc base c nhm i, tc 2 nucleotid m ho cho mt loi acid amin th
tt c ch c 16 t hp, khng cho 20 loi acid amin. Nh vy, n v m ho hay cn gi l codon
phi gm 3 hay nhiu nucleotid hn.
Nm 1961, F. Crick lm th nghim chng minh rng nhm nucleotid m ho c 3 hay ni cch
khc codon gm 3 nucleotid k tip nhau. Tt c s c 43 = 64 t hp.
M. W Nirenberg v H. Matthaei (M) dng enzym theo phng php ca Ochoa tng hp ARN
nhn to. Khi ch dng mt loi nucleotid l Uracil s nhn c ARN l polyuracil, nu ch Adenin s
nhn c polyadenin.
Nm 1961, khi dng polyuracil thay cho mARN tng hp protein trong h thng v bo (c acid
amin, enzym, nhng khng c ADN...), sn phm nhn c l mch polypeptid polyphenylalanin. iu
chng t codon UUU m ho cho phenylalanin. y l codon u tin c xc nh. Nirenberg v
Matthaei cng chng minh c AAA m cho lysin, GGG cho Glycin, CCC cho prolin.
Nm 1964, H. G. Khorana tm ra phng php to mARN nhn to vi trnh t lp li (nh

AAGAAGAAG...) v nh gii quyt cc vn cha r.
Bng m di truyn (bng 4.3) cho thy trong 64 codon c 3 codon UAA, UAG, UGA khng m ho
cho acid amin c gi l codon v ngha (non - sense), ng thi l codon kt thc (stop) tc l "du
chm cu", chm dt mch polypeptid.
M di truyn c tnh "suy thoi" tc mt acid amin c nhiu codon m ho, ch tr methionin v
tryptophan ch c mt codon.
Cc codon ng ngha tc l m ho cho cng mt acid amin thng c 2 base u tin ging nhau,
nhng khc nhau nucleotid th 3. V d: CCU, CCC, CCA v CCG u m ho cho prolin. Trn thc
t U v C lun lun tng ng nhau v tr th 3, cn A v G tng ng nhau trong 14 ca 16 trng
hp.
Tr mt s ngoi l, m di truyn c tnh ph bin cho tt c sinh vt.
Mt codon ch m cho mt loi acid amin, trng hp ngoi l l AUG va m ho cho Met ni,
va m ho cho acid amin m u N - formyl Methionin trong t bo t bo nhn nguyn thu hoc
methionin trong t bo nhn tht.
Bng 4.3. M di truyn gm 64 codon
V tr th II
V tr th I U C A G V tr th III
Phe Ser Tyr Cys U
Phe Ser Tyr Cys C

U
Leu Ser stop stop A
Leu Ser stop Trp G
Leu Pro His Arg U
Leu Pro His Arg C

C
Leu Pro Gln Arg A
Leu Pro Gln Arg G
Ile Thr Asn Ser U
Ile Thr Asn Ser C

A
Ile Thr Lys Arg A
Met Thr Lys Arg G
Val Ala Asp Gly U
Val Ala Asp Gly C

G
Val Ala Glu Gly A
Val Ala Glu Gly G
Tm tt
Qu trnh phin m trong t bo nhn nguyn thu v t bo nhn tht c mt vi chi tit khc nhau
ng k. T bo nhn nguyn thu ch cha mt loi enzym nhn ARN polymerase, m khi tng tc
vi mt tiu n v iu ho (sigma) s to ra holoenzym quy nh tnh c hiu ca promoter. Promoter
gm 2 vng cha cc trnh t nm trung tm - 10 v - 35 k t v tr bt u phin m. Cc trnh t ny
tng tc vi holoenzym ARN polymerase lm cho s phin m c hiu qu v chnh xc.
Nhn t bo nhn tht cha 3 loi ARN polymerase khc nhau. Polymerase I phin m cc rARN;
polymerase II phin m cc gen m ho protein; polymerase III phin m rARN 5S, tARN v cc ARN
nh khc. Cc promoter ca nhng nhm gen trn c khc bit ng k. Cc gen c phin m bi
polymerase II c trnh t nhn bit ngay trong cc promoter ca chng, nhng cng c nhiu s khc
bit. Cc promoter thuc ARN polymerase III khc thng l nm ngay trong trnh t ca gen c
phin m.
Khc vi cc promoter ca t bo nhn nguyn thu, cc promoter t bo nhn tht khng l cc v

tr gn trc tip ARN polymerase, m thc hin chc nng kt dnh cc yu t phin m (protein) lm
cho nhim sc th tho xon, cho php ton b ARN polymerase xm nhp vo. Ngoi ra, nhiu gen t
bo nhn tht cn c hot ho bi cc yu t gn vo trnh t tng cng (enhancer element) trc
khi phin m c hiu qu.
M di truyn gm b 3 cc base (codon) trong mARN. Chng chuyn bit cho vic kt hp acid
amin vo mch polypeptid. M c tnh theo chiu 5 3 ca mARN, lin tc, ph qut v suy thoi,
trong nhiu loi acid amin c quy nh bi 2 hoc nhiu hn 1 codon.
CU HI
1. Phin m i xng c th xy ra :
a) Vi khun b) Lp th
c) Ti th ca rui gim d) Ti th cn trng
e) Ti th t bo th
2. Tiu n v tch ra khi phc hp phin m khi ARN mi sinh t chiu di:
a) 4 base b) 6 base
c) 8 base d) 10 base
e) 12 base
3. V sao ARN polymerase khng cn c hot tnh sa sai?
a) Nucleotid kt hp khng ng c thay th ngay
b) Sai st him hoi khng di truyn c
c) ARN khng phi l ni lu tr thng tin di truyn
d) ARN khng to ra chnh n
e) a, b, c
4. ARN polymerase prokaryote l mt holoenzym cha cc tiu n v:
a) b) '
c) ' d) ''
e)
5. E. coli, promoter gm cc vng:
a) Vng TATAAT b) Hp TATA
c) Vng TTGACA d) Vng 35
e) Tt c
6. Chc nng quan trng ca hp 10 v hp 35 c pht hin nh t bin:
a) Mt base
b) Thay base ny bng base khc
c) Thm base
d) o v tr mt cp base
e) a, c
7. S kin khng ng vi hin tng phin m ngc:
a) Cn mi
b) on mi l tARN ca t bo ch
c) on mi l ARN do primase tng hp
d) on mi gn vo u 3 ca Retrovirus
e) ARN virus trong chui lai b phn hu bi RNaseH
8. c im no khng thuc s phin m t bo nhn tht?
a) mARN cha thng tin mt gen
b) u 5 mARN c gn chp 7 - Methylguanosine
c) Bn phin m u tin (pre - mARN) c s dng ngay cho vic tng hp protein
d) C thm ui polyA di 100 - 200 nucleotid
e) C 3 loi ARN polymerase I, II v III
9. Acid amin no ch c mt codon?
a) Leucin b) Methionin
c) Tryptophan d) Alanin
e) b v c
10. Tnh cht no khng phi ca m di truyn?
a) C ngoi l
b) Mt chiu, khng chng ln nhau
c) Ph bin mi sinh vt l m b 3
d) c hiu, mt codon ch m ho cho mt loi acid amin
e) Suy thoi, nhiu b ba m ho cho mt loi acid amin
Bi 5
SINH TNG HP PROTEIN
MC TIU
- M t c c ch dch m di truyn.
- Phn bit c qu trnh dch m t bo nhn tht v t bo nhn nguyn thu.
- Da vo c ch khc nhau ny gii thch c tc ng chn lc ca khng sinh.
- M t c cch thc m bo s chnh xc ca qu trnh tng hp protein.
5.1. M U
Giai on cui ca s biu hin thng tin di truyn xy ra khi thng tin c m ho bng trnh t
nucleotid trn mARN c dch m thnh trnh t acid amin tng ng trong chui polypeptid. Qu
trnh ny c gi l qu trnh tng hp protein. Mc d, v c bn tin trnh dch m xy ra nh nhau
mi sinh vt, nhng vn c nhng chi tit khc nhau gia t bo vi khun v t bo nhn tht. Nhn
chung, qu trnh ny t bo nhn tht phc tp hn. Phn ln qu trnh tng hp protein xy ra trong t
bo cht. Mt s bo quan ca t bo nhn tht cng c b my di truyn ring (lc lp, ty th) v cng
c kh nng tng hp c protein m ho bi chng. B my dch m ca cc bo quan ny ging t
bo nhn nguyn thu hn l t bo nhn tht.
5.2. CC YU T CN THIT CHO S TNG HP PROTEIN
5.2.1. Ribosom
Ribosom l xng tng hp protein ca t bo. Tuy c s khc nhau trong cu to v kch thc
gia cc gii sinh vt, nhng u gm 2 tiu n v. V ch c cc ribosom hon chnh, gm mt tiu
n v ln v mt tiu n v nh mi c kh nng dch m mARN.
Khi khng thc hin tng hp protein, mi tiu n v tn ti tch ri trong t bo cht (hnh 5.1).
Tiu n v ln ca t bo nhn nguyn thu gm 2 phn t rARN v 35 phn t protein, tiu n v
nh c 1 phn t rARN v khong 20 phn t protein. rARN c cu trc khng gian phc tp do nhiu
on bt cp vi nhau nh c trnh t nucleotid b sung.
Hnh 5.1. Cc tiu n v ribosom v s dch m

(a) 2 tiu n v ribosom vi mARN; (b) s dch m ca ribosom
t bo nhn nguyn thu, u 5 ca mARN gn trc tin vo tiu n v nh v lc c kh

nng gn thm vo tiu n v ln. Khi n v ln gn xong th s dch m bt u.
V t bo nhn nguyn thu khng c mng nhn, qu trnh phin m din ra trong t bo cht, cc
ribosom c kh nng gn vo mARN ang c phin m v dch m to ra protein trong khi ARN
polymerase vn tip tc phin m t ADN.
c t bo nhn nguyn thu ln t bo nhn tht, khi ribosom u tin dch m mARN c mt
on, th cc ribosom khc c th gn vo pha trc dch m. Do , cc ribosom xp thnh chui
trn mARN to nn cu trc polyribosom hay cn gi l polysome (hnh 5.2). Khong 15 - 20 ribosom
c th gn cng lc trn mARN. Nh vy, tc tng hp protein tng nhanh ng k.
Hnh 5.2. Polyribosom
5.2.2. ARN thng tin
Cu trc ca mARN in hnh c minh ha hnh 5.3. Thng tin di truyn l mt trnh t thng
ca cc base nucleotid. Trnh t trong vng m ho ca mARN cha cc codon quy nh trnh t acid
amin ca protein. Trnh t ny c c theo hng 5 n 3 trn mARN c m u bng mt codon
m u (thng l AUG). Kt thc l mt trong cc codon kt thc (UAA, UGA hoc UAG).
Phn t mARN chm dt ti mt trnh t khng m ho v cng c mt vng khng m ho u

5 c gi l trnh t dn bao gm mt trnh t gn ribosom t bo nhn nguyn thu. Cc trnh t
dn u 5 rt khc nhau v di trong cc loi mARN khc nhau, nhng nhn chung, t bo nhn
tht u di hn t bo nhn nguyn thu. Cc trnh t 3 v 5 khng m ho cn c gi l cc
vng khng dch m (UTR).
Ngoi cc trnh t khng m ho u 5 v 3, cc mARN t bo cht t bo nhn tht c gn thm

"chp" (CAP) 7 - methyl Guanosine Triphosphat u 5 (hnh 5.4). c im ny lin quan n s
tng tc gia mARN v ribosom. Cc trnh t khng dch m u tn cng 3 c chiu di rt khc
nhau. Hu ht cc phn t mARN t bo nhn tht u 3 c thm ui 100 - 200 Adenin sau khi
phin m. Chc nng ca ui poly(A) cha c r rng. Chng ng vai tr quan trng trong vic bo
v mARN khi b thu gii ca exonuclease, nhng li khng cn thit cho s dch m. Mt vi loi
mARN nh histon - mARN v mARN vi khun khng c ui poly(A).
Hnh 5.3. Cc cu trc chi tit ca cc phn t mARN

(a) mARN vi khun c cha 2 cistron; (b) monocistron mARN t bo nhn tht
Hnh 5.4. Cu trc chp (CAP) ca t bo nhn tht
5.2.3. ARN vn chuyn v cc enzym tng hp tARN - aminoacyl
Cc ARN vn chuyn hot ng nh mt cht kt ni gia thng tin base - nucleotid ca mARN vi
trnh t acid amin ca phn t protein tng ng. Mi tARN c hiu cho mt loi acid amin ring bit,
mc d n c th gii m cho hn mt codon, s lng cc tARN bin ng theo loi: t bo nhn
nguyn thu 30 - 40, t bo nhn tht 50. Do , cc tARN phi c nhn bit chuyn bit ng thi
bi mARN v bi enzym gn acid amin vo tARN tng ng (enzym tARN - aminoacyl synthetase).
iu ny gip gii quyt tr ngi khng gian trong qu trnh dch m. Kch thc ca codon ln hn
nhiu so vi acid amin, nn nu mt acid amin nhn bit v gn trc tip ln mt codon trn mARN th
n s cch xa vi acid amin tng ng vi codon k cn hnh thnh lin kt peptid. V c ti 20 loi
acid amin khc nhau cu trc nn bn dch, do , cn phi c ti 20 loi aminoacyl - tARN synthetase
khc nhau. l Alanyl - tARN synthetase, Arginyl - tARN synthetase, Asparaginyl - tARN
synthetase,... Mc d, c th c ti mt vi loi tARN khc nhau cho mi mt loi acid amin ( gii m
cc codon khc nhau tng ng vi acid amin ), nhng ch c mt aminoacyl - tARN synthetase cho
mi loi acid amin. Vn mt aminoacyl - tARN synthetase nhn din c vi tARN khc nhau nh
th no hin vn cha r rng. Dng nh, s nhn din chnh xc tARN ca synthetase thng qua vic
nhn din anticodon. Enzym aminoacyl - tARN synthetase phi ph hp v chn ng tARN: tARN ch
ng cho mt synthetase th c gi l tARN duy nht (cognate tARN). Cc loi tARN lin quan n
cng mt acid amin th c gi l tARN ng nhn (isoaccepting tARN).
Vn mu cht l cc bc aminoacyl ho phi chnh xc. Bi v, nu mt tARN chp nhn mt

acid amin sai dn n vic gn kt khng ng vo protein v s khng c "bc c sa sai" theo sau
s aminoacyl ho. Bc c sa sai ch c thc hin giai on aminoacyl ho bi chnh enzym
aminoacyl - tARN synthetase. Qu trnh aminoacyl ho cng lc thc hin 2 chc nng. Trc tin, n
cung cp s lin h u tin ca m di truyn (da vo acid nucleic) v cu trc protein (vi cc acid
amin). Tip n, n hot ho acid amin trc khi kt ni vo protein. Lin kt ester aminoacyl gia acid
amin v tARN d tr mt nng lng thu gii t do tng i cao, nng lng ny rt cn thit cho
vic hnh thnh lin kt peptid trong qu trnh dch m.
5.2.4. Aminoacyl ho
Qu trnh aminoacyl ho xy ra theo 2 bc:
5.2.5.1. Hnh thnh aminoacyl - adenylate hot ho
Acid amin phn ng vi ATP c enzym xc tc to thnh aminoacyl adenylate hot ho.
Pyrophosphat to ra b thu gii thnh 2 phn t phosphat v c. Phn ng ny to ra mt nng lng
thu gii t do tng i cao, y phn ng theo chiu to aminoacyl - AMP trung gian:
5.2.5.2. Hnh thnh aminoacyl - tARN
Aminoacyl - AMP c chuyn ti phn t tARN tng ng bng synthetase:
C hn mt codon trong m di truyn cho tng loi acid amin, ngoi tr methionin v tryptophan.
Nhng codon ng ngha (synonym) ny thng lin quan n cc tARN khc nhau. Cc tARN li c
trnh t i m (anticodon) khc nhau. Th d, trong trng hp ca serin, mc d c ti 6 codon (UCU,
UCC, UCG, UCA, AGU, AGC) u c ngha nh nhau, nhng khng phi tt c u c s dng
thng xuyn nh nhau. Tht vy, c mt vi codon cho mt acid amin c s dng v c mt vi
phn t tARN cho mi codon. Hin tng ny c gi l s d bit codon v y l im khc nhau
quan trng nht gia loi ny v loi khc.
5.2.5. Cc tARN v cc codon khi u
Trnh t m ca phn t mARN c khi u vi codon AUG, mc d vi khun cn c codon

GUG c s dng cho mt s mARN (khong 4% trnh t c GUG khi u c bit). Rt him
khi cc codon khc (AUU, UUG) hot ng nh codon khi u.
Du hiu khi u l codon AUG, cng l codon m ho cho methionin (Met). Mc d, ch c mt

codon m ho cho Met, nhng trong t bo cht li hin din 2 loi tARN c kh nng mang Met n
kt hp vi codon t bo nhn nguyn thu:
(1) tARNmMet kt hp vi codon AUG nm gia phn t mARN, c nhim v gn Met vo chui
polypeptid ang hnh thnh.
(2) tARNiMet kt hp vi codon AUG khi ng, gn Met m u vo chui polypeptid.
vi khun, tARNiMet khi u khng ch vn chuyn methionin m cn vn chuyn formyl -

methionin: tARN ny c gi l tARNfMet (vit tt l fMet) (hnh 5.5).
Hnh 5.5. Formyl - methionyl - tARN
S tng hp tt c protein vi khun u bt u bng formyl - methionin, mc d nhm formyl,

cng nh nhm methionyl, thng b loi b sau khi tng hp xong phn t protein. V d, E. coli, ch
c 45% cc protein hon chnh bt u bng methionin. Mc d codon khi u khng phi AUG (GUG
chng hn) vn c gii m bi fMet - tARNfMet.Cn phn t tARN lin quan ti codon AUG bn
trong mARN lm nhim v vn chuyn methionin c vit tt l tARNmMet.
t bo nhn tht, cc tARN chuyn bit cho cc codon AUG khi u v AUG bn trong mARN,
nhng tARNMet m u ch vn chuyn methionin khi u c gi l tARNiMet v tARNmMet cho
cc codon AUG bn trong (bng 5.1).
Bng 5.1. Cc codon khi u v tARN khi u
Vi khun T bo nhn tht
Codon khi u AUG (GUG) AUG
tARN khi u tARNfMet tARNIMet
tARN mang Formyl - methionin Methionin
5.3. DIN BIN DCH M RIBOSOM (CHU TRNH RIBOSOM)
Qu trnh dch m mARN c m t trong hnh 5.6, bao gm cc giai on: khi u, ni di v
kt thc.
5.3.1. Khi u
Giai on khi u, c s tham gia ca 2 tiu n v ca ribosom, mt phn t mARN (c sp

thng hng vi ribosom) v tARN khi u mang methionin hoc formyl - methionin t bo nhn
nguyn thu (bng 5.2). V cc yu t khi u nh IF1, IF2, IF3 vi khun v 10 yu t eIF (IF ca t
bo nhn tht) nh eIF1, eIF2, eIF3, eIF - 4A, eIF - 4B... t bo nhn tht (bng 5.2).
Hnh 5.6. Chu trnh ribosom t bo nhn tht
5.3.1.1. Khi u chui peptid vi khun
c trng ca s khi u chui peptid vi khun l s tng tc trc tip gia mARN v rARN
ca tiu n v nh 30S (hnh 5.7). S tng tc ny dn n vic gn mARN vo ribosom v quan
trng hn l nhn din c codon khi u ca trnh t mARN. Cc mARN vi khun c cha mt trnh
t dn u 5 kh ngn. N bao gm trnh t polypurin ngn b sung vi mt trnh t giu pyrimidin
tn cng 3 ca rARN 16S (hnh 5.8). Shine v Dalgarno cho rng vic ghp i base gia cc trnh t
ny c th l c ch lm cho ribosom ca vi khun hng ti ng codon khi u ca mARN. Do ,
ngi ta gi trnh t ngn ny l trnh t Shine - Dalgarno.
Hnh 5.7. S khi u dch m vi khun
Hnh 5.8. Vai tr ca trnh t Shine Dalgarno
Vai tr IF - 2 l lm trung gian trong vic ghp tARN khi u (fMet - tARNfMet) vo tiu n v
30S. iu ny ph thuc vo GTP. Bi v, IF - 2 to ra phc 3 yu t vi GTP v vi tARN khi u.
l phc [IF - 2 - GTP - fMet - tARNfMet] m tARN gn vi ribosom. Cc phn t tARN lun
lun gn vi ribosom nh nhng phc c cha GTP v yu t protein. Chc nng chnh xc ca IF - 1
cha c xc nh, nhng trong mt s trng hp cho thy n to iu kin cho tc ng ca IF - 2.
C th mARN hoc phc hp tARN gn trc tin vo tiu n v 30S. Ngi ta mi ch bit c
rng, c 2 bc ny u xy ra trc khi tiu n v 50S gn vo to ra phc hp 70S cho vic ni
di. Vai tr ch yu ca IF - 3 l gi s gn kt mARN vi ribosom.
S khi u chui peptid vi khun bao gm s nhn din trnh t Shine - Dalgarno, nm gn im
xut pht ca trnh t m ho mARN. Do , s khi u dch m khng nht thit nm gn vi im
khi u 5 ca mARN. ng vy, cc mARN t bo nhn nguyn thu c cha ti vi vng m ho,
ng hn l vi khun thng c polycistron mARN. c bit, r nht l mARN cha operon m ho
cho 3 loi protein khc nhau lac operon.
5.3.1.2. Khi u chui peptid t bo nhn tht
Qu trnh khi u chui peptid t bo nhn tht phc tp hn v c nhiu im khc bit vi t
bo nhn nguyn thu. Trong khi, E. coli ch c 3 yu t khi u th t bo nhn tht c ti 10.
iu ny c phn nh trong cc cch thc gn kt mARN v xc nh codon khi u.
Trnh t dn mARN t bo nhn tht khng c xem nh trnh t Shine - Dalgarno t bo nhn
nguyn thu. u tn cng 3 ca rARN 18S (tng ng vi rARN 16S vi khun) khng c trnh t
CCUCC. V mARN t bo nhn tht l monocistron, chng ch cha mt trnh t m ho v khi u,
thng xy ra codon khi u AUG trong mARN. Nhng nhn xt trn gip cho Kozak xy dng m
hnh khi u sinh tng hp protein t bo nhn tht (hnh 5.9).
Trc tin tiu n v nh 40S ca ribosom gn vo u tn cng 5 ca mARN, sau di chuyn

dc theo mARN ti khi gp codon khi u AUG. Nng lng cho s di chuyn ly t s thu phn
ATP v s tng tc khi u vi mARN c th lin quan ti cu trc chp 5 (CAP). 95% cc trng
hp khi u t bo nhn tht xy ra ti codon khi u AUG, hu nh khng dng ti GUG.
(a) Tiu n v nh 40S gn vo mARN chp 5 v qua chp (Cap) hnh thnh nn phc hp chp
- cc yu t khi u.
(b) Tiu n v 40S qut dc theo mARN, tm AUG khi u. Qu trnh ny cn ATP v c th tho
xon nt kp tc.
(c) Codon khi u, thng l AUG c la chn.
Hnh 5.9. M hnh Kozak
Phc hp mi hnh thnh by gi c th tng tc vi tiu n v 60S tham gia vo giai on

ni di.
Codon AUG c th ch l codon m ho bnh thng nhng cng c th l codon khi u, nu n

c nh v trong mt "ng cnh" ng. Ng cnh bit ti nhiu nht c trnh by hnh 5.10.
Hnh 5.10. Ng cnh nhn din codon khi u

Nhng nucleotid vit m l nhng nucleotid quan trng trong trnh t chung.
Tiu n v 40S s b qua codon AUG bnh thng cho n khi nh v ti codon AUG khi u
nm trong ng cnh tt.
Bng 5.2. Cc yu t khi u ca t bo nhn nguyn thu

v t bo nhn tht v chc nng ca chng
Yu t t bo
Yu t vi khun Chc nng
nhn tht
IF - 1 eIF - 1 Vai tr cha r
IF - 2 eIF - 2 Gn Met - tARN (hoc fMet - tARN) vo ribosom

thnh phc hp vi GTP
IF - 3 eIF - 3 Ngn cn s tch i ca cc tiu n v ribosom
eIF - 4A Nhm yu t lin quan p ng cho vic gn chp

5 vo u mARN v tho xon cu trc bc 2 to
eIF - 4B iu kin cho ribosom u vo codon khi u
eIF - 4E
eIF - 4F
eIF - 5 Gii phng eIF - 2 v eIF - 3 khi ribosom v gip

cho tiu n v 60S kt ni
eIF - 6 Tham gia vo vic tch ribosom thnh cc tiu n

v
GEF Yu t bin i nucleotid Guanin: ti sinh eIF - 2 bi

s bin i trung gian GDP thnh GTP trong qu
trnh tng t ti sinh EF - Tu bi EF - Ts
(hnh 5.7)
Bng 5.2 gii thiu 10 yu t khi u ca t bo nhn tht. y, eIF - 3 (ging IF - 3) cn tr

cc tiu n v ribosom tch i. eIF - 2 (ging IF - 2) lm trung gian gn kt tARN - khi u vi tiu
n v nh thnh phc hp vi GTP. Tuy nhin, t bo nhn tht Met - tARNIMet phi c gn kt
trc vo mARN. iu cho thy rng i m (CAU) trn tARN - khi u nhn ra AUG trong khi
tiu n v 40S ang qut trn mARN.
eIF - 6 tng tc vi tiu n v ln 60S v tch i c 2 tiu n v ca ribosom. eIF - 5 gip

tiu n v 60S kt ni v gii phng cc yu t khi u khc v cc yu t khc lin quan ti s gn
mARN nh eIF - 4A, eIF - 4B, eIF - 4E, eIF - 4F.
Cc yu t ny thng l nhng protein gn chp do chng gn vo chp 5: chng gn vo cc hp

phn ca chp (nh f - methyl GTP chng hn). eIF - 4F l mt phc hp protein cha cc thnh phn
tng t eIF - 4E v eIF - 4A, v cng l protein c phn t lng Mr 220000. Ngi ta cho rng eIF -
4F thng qua eIF - 4E gn trc tip vo chp trc v sau eIF - 4A v eIF - 4B mi gn vo.
Cc trnh t dn u 5 ca mARN t bo nhn tht thng di n vi trm base v c th cha cc

vng b sung, lm nh ln cc nt kp tc. y l cu trc bc 2 ca mARN. cho ribosom c th
dch m c, iu cn thit l phi tho xon cu trc bc 2 ny. Bi v, c th cu trc bc 2 ny ngn
cn s nhn dng ca ribosom i vi mARN hoc che khut codon khi u AUG.
eIF - 4A c kh nng tho xon cc cu trc bc 2 (cn thu phn ATP). Vai tr ca eIF - 4B cha
r, nhng c th lm gia tng tc ng ca eIF - 4A (hnh 5.11).
Hnh 5.11. S khi u t bo nhn tht
5.3.2. Ni di
Qu trnh dch m trn mARN c ni di theo hng 5 3 v chui polypeptid c bt u

tng hp u tn cng - N. Qu trnh ny ging nhau t bo nhn nguyn thu v t bo nhn tht
(hnh 5.13).
S ni di l mt qu trnh c chu k, m mi chu k chu trch nhim gii m mt codon v thm

mt acid amin vo chui polypeptid mi sinh. S ni di cn n cc protein, c gi l cc yu t ni
di (EF). vi khun, cc yu t ny l EF - Tu, EF - Ts v EF - G cn t bo nhn tht l EF - 1
(tng ng EF - Tu) v EF - 2 (tng ng EF - G), EF - 1 - Tu (tng ng EF - Ts (bng 5.3).
Trn ribosom c 2 v tr A v P. V tr A l v tr Aminocyl (hoc Acceptor) v v tr P l v tr
Peptidyl, ni gi phc hp peptidyl - tARN, tc l chui polypeptid ang hnh thnh vn cn gn

vi tARN trc .
Bng 5.3. Cc yu t ni di
t bo nhn
t bo nhn tht Chc nng
nguyn thu
EF - Tu EF - 1 Mang cc aminoacyl - tARN ti v tr A ca

ribosom, th d, phc hp (EF - 1 - GTP -
(Mr = 43 000) (Mr = 53 000) aminoacyl - tARN).
EF - Ts EF - 1 - Tu Ti hi EF - Tu hoc EF - 1 (hnh 5.14).
(Mr = 30 000) (Mr = 50/38 000)
EF - G EF - 2 Tham gia vo bc chuyn v ca chu k ni

di.
(Mr = 77 000) (Mr = 100 000)
im khi u mi chu k ni di, ch c v tr P b chim ch bng mt peptidyl - tARN cha

chui polypeptid mi sinh gn vi tARN mang acid amin va c thm vo. Peptid c gn vo
tARN theo cng mt cch nh acid amin, bng nhm 3 - hydroxyl v tr kt thc CCA ca tARN
(hnh 5.12).
V tr P c ging hng vi codon k sau c dch m, trong khi v tr A ging hng vi

codon k tip ca mARN. Lc ny v tr A trng. Bt k aminoacyl - tARN (ngoi tr tARN - khi u)
u vo v tr A, chng lu li y nu anticodon khp m vi codon ti A. Bng cch ny, aminoacyl -
tARN chnh xc c la chn v chn c ng acid amin cho s hnh thnh chui polypeptid mi
sinh. Tht ra, vic gn ny l gn phc hp gia EF - Tu hoc EF - 1 vi GTP ging nh tARN - khi
u gn kt trong sut giai on khi u.
Hnh 5.12. Cu trc ribosom
Hnh 5.13. Qu trnh ni di gn nh ging nhau gia vi khun v t bo nhn tht
Theo sau s gn kt phc hp vo v tr A, l qu trnh thu phn GTP gii phng GDP v Pi (hnh
5.13). GDP c phng thch khi ribosom s gn kt vi yu t ni di to phc hp [EF - Tu - GDP]
hoc [EF - 1 - GDP]. yu t ni di li c th tham gia vo qu trnh gn kt aminoacyl - tARN tip
theo vo ribosom th GDP phi c thay th bi GTP. vi khun, yu t EF - Ts ng vai tr trung
gian cho vic chuyn i GDP/GTP (hnh 5.14), tng t cc yu t ca protein - G lin quan ti s ti
np tn hiu hormon. Gia acid amin sau cng ca chui mi sinh vi acid amin trn tARN v tr A,
hnh thnh lin kt peptid. Phn ng ny c xc tc bi enzym peptidyl - transferase. Kt qu l
tARN v tr P lc by gi khng cn mang acid amin hoc l tch khi chui polypeptid.
Bc chuyn v (translocation step) cui cng ca giai on ni di cn mt yu t ni di khc l

EF - G hoc EF - 2 v GTP. GTP thu phn thnh GDP v Pi. S chuyn v c 3 hin tng xy ra ng
thi:
- tARN vn chuyn xong c tch khi v tr P
- Peptidyl - tARN di chuyn t v tr A sang v tr P
- Ribosom chuyn dch theo tng codon trn mARN, do , codon tip theo c a vo ging
hng vi v tr A.
Qu trnh chuyn v ni di s c lp li cho n khi c mt codon kt thc xut hin v tr A.

Vai tr ca GTP trong qu trnh ni di cha r rng. Ngi ta cho rng s thu phn GTP cung cp
nng lng cho vic to lin kt peptid v cho bc chuyn v, nhng n c th lin quan n mc
chnh xc ca s chuyn v.
Hnh 5.14. S ti hi EF - Tu bi EF - Ts v ng phn ca protein - G
EF - Tu c gii phng khi ribosom sau mi chu k ni di thnh phc hp vi GDP (bt hot),
ngha l phc hp ny khng th gn kt aminoacyl - tARN. cho EF - Tu tham gia vo vng ni di
tip theo th GDP cn phi c bin i thnh GTP to phc hp c hot tnh [EF - Tu - GTP]. S bin
i nucleotid Guanin cn c mt yu t protein khc, l EF - Ts.
5.3.3. Kt thc
Chu trnh dch m thc hin thm khong 15 acid amin mt giy vo mch polypeptid. Chu trnh
c chm dt khi tri qua codon kt thc l UAA, UAG v UGA.
bc kt thc, cc m kt thc khng c anticodon. Thay vo l cc yu t kt thc (RF) lm

kt thc qu trnh dch m.
vi khun c 3 yu t phng thch: RF - 1 chuyn bit cho cc codon UAA v UAG; RF - 2 lin
quan ti UAA hoc UGA v RF - 3 kch thch c 2 yu t kia. y cn cn yu t ni di EF - G v
yu t phng thch ribosom (RRF) chuyn bit.
t bo nhn tht ch c mt yu t phng thch eRF c bit (bng 5.4).
Bng 5.4. Cc yu t kt thc
Vi khun (E. coli) T bo nhn tht
RF - 1 (cho UAA hoc UAG)
RF - 2 (cho UAA hoc UGA) eRF (ph hp vi c 3 codon kt thc)
RF - 3 (Kch thch RF - 2)
S kt thc lin quan n vic phn gii lin kt gia acid amin cui cng ca chui peptid mi sinh
v tARN cui cng. Phn ng ny c xc tc bi enzym peptidyl transferase ca ribosom v i hi
GTP. Nh , polypeptid hon chnh c phng thch. Sau , 2 tiu n v ca ribosom c phng
thch cng vi mARN. Tt c cc thnh phn tham gia chu trnh u c s dng li.
5.4. NHU CU NNG LNG CHO QU TRNH SINH TNG HP PROTEIN
mt vi giai on ca qu trnh sinh tng hp protein cn n nng lng chuyn ho v nng

lng ny c ly t s thu phn ATP v GTP (bng 5.5).
Bng 5.5. Nhu cu nng lng cho qu trnh dch m
Giai on T bo nhn nguyn thu T bo nhn tht
Aminoacyl ATP AMP + PPi ATP AMP + PPi

ho tARN
(2 phosphoanhydrid
cho mt gc acid)
Khi u GTP GDP + Pi, GTP GDP + Pi,

kt hp vi IF - 2 kt hp vi eIF - 2
ATP ADP + Pi,

kt hp vi vic gn chp v tho xon
mARN (cha r c bao nhiu phn t
ATP c s dng)
Ni di GTP GDP + Pi, GTP GDP + Pi,

kt hp vi EF - Tu kt hp vi EF - 1
GTP GDP + Pi, GTP GDP + Pi,

kt hp vi EF - G kt hp vi EF - 2
Kt thc Khng cn nng lng GTP GDP + Pi
Nng lng cn cho cc giai on khc nhau, c th ng mt hoc mt s vai tr sau:
- Cung cp nng lng thc hin cc phn ng c bit, nh hot ho acid amin bng cch gn
chng vo tARN.
- Cung cp nng lng cho mt qu trnh no nh tho xon cu trc bc 2 ca mARN hoc cho
s chuyn dch ribosom trong cc bc chuyn v trong qu trnh ni di.
- Duy tr cc yu t trng thi hot tnh nh EF - Tu.
- Cung cp nng lng cho s bin dng ca ribosom, nh sau khi gn kt aminoacyl - tARN vo v
tr A.
C th tnh nng lng chuyn ho cn dng cho s tng hp mt chui polypeptid in hnh vi
khun gm 300 acid amin nh sau:
- aminoacyl ho tARN cn 300ATP 300AMP 600 lin kt phosphoanhydrid.
Mt phn ng khi u, 1GTP 1GDP 1 lin kt phosphoanhydrid
Cho 229 vng ni di:
299 GTP 299 GDP giai on EF - Tu 299 lin kt phosphoanhydrid
299 GTP 299 GDP giai on EF - G 299 lin kt phosphoanhydrid
Tng nng lng cn 1198 lin kt phosphoanhydrid
Tm li, tnh ra chnh xc c 4 lin kt phosphoanhydrid c s dng cho mi acid amin trong qu
trnh sinh tng hp protein.
5.5. CHNH XC CA QU TRNH DCH M
Sai st trong qu trnh dch m s to ra mt protein bt hot, hoc l mt protein iu ho v xc

tc khng ph hp lm thay i sinh l ca t bo bnh thng. Cc sai st l:
5.5.1. Aminoacyl ho tARN
Mt tARN c th nhn thm mt acid amin khng ng hoc enzym aminoacyl - tARN synthetase
dng tARN c anticodon khng chuyn bit cho cc acid amin c gn vo n. C mt s acid amin c
cu trc tng t vi acid amin cn c a vo, cho nn cc enzym aminoacyl - tARN synthetase
cng phi c cch thc chn ng.V d, enzym isoleucyl - tARN synthetase, mt mt phi phn bit
gia isoleucin v cc acid amin nh hn nh valin, mt khc phi phn bit gia isoleucin vi cc acid
amin k nc, cng knh hn nh leucin, phenylalanin. C ch iu chnh xy ra nh sau:
Enzym isoleucyl - tARN synthetase c 2 v tr tc ng, mt v tr xc tc thnh lp aminoacyl -

tARN (A) v v tr th 2 (H), st ngay v tr th nht, thu phn aminoacyl - tARN cho ra acid amin
t do v tARN. Cc acid amin ln hn isoleucin (hoc c cu hnh khng ng) khng th i vo v tr
acyl ho (A) v do khng c cc enzym synthetase s dng. Tuy nhin, cc acid amin nh hn
isoleucin c th i vo v tr A to ra aminoacyl - tARNile v s l ngun gc tim tng gy ra cc sai
st. Cc dn xut aminoacyl cng c th i vo v tr thu phn (H), nn b thu phn. V vy, ch c
isoleucyl - tARN l c sn sinh chuyn bit. 2 bc nhn bit trn lm tng chnh xc ca s
aminoacyl ho tARN. Bc sa sai c minh ha bng hnh 5.15.
5.5.2. Tng tc codon - anticodon
Phc hp sai gia codon v anticodon s dn n vic gn vo acid amin khng ng. Cc nghin
cu t bin vi khun cho thy cc protein chuyn bit trong cc tiu n v ca ribosom ng vai tr
quan trng trong vic gn ng cc codon v anticodon trong sut qu trnh dch m.
5.5.3. Kt thc sm, kt thc mun
Cc sai st ca giai on kt thc c th xy ra do kt thc sm (premature termination) hoc kt

thc mun do c qu. S c qu do li ca b my dch m, khng nhn ra codon dng. Tt nhin,
acid amin c a vo v dch m lin tc qua c vng khng m ho 3mARN. S c qu cng c
th do mt t bin codon i m no lm cho codon i m ny gii m c codon kt thc.
(a) Cc acid amin ln hn isoleucin khng vo c v tr A

(nh phenylalanin). Kt qu: phn ng khng xy ra
(b) Cc acid amin nh hn isoleucin (nh valin) vo v tr A

v to aminoacyl-tARN, nhng cng i vo c v tr H nn b
thu phn thnh 2 phn tARN v acid amin. Kt qu: phn ng
khng xy ra
c) Ch c isoleucin gn vo v tr A to isoleucyl-tARN
nhng khng th gn vo v tr H v do khng b thu phn.
Kt qu: Acyl ho chnh xc isoleucin
Hnh 5.15. S bc sa sai aminoacyl ho tARN ca enzym synthetase

A: v tr acyl ho (acylation); H: v tr thu phn (hydrolytic).
Ngi ta c tnh rng, sai st tng ln t l 1/3000 gc acid amin c kt hp. Ngha l, nu mt
phn t protein c 300 gc acid amin th ch 1 phn t protein trong 10 phn t c cha 1 li v thng
nhng li ny s c nh v m khng quan trng i vi cu trc hoc chc nng ca protein.
5.6. CC YU T C CH QU TRNH DCH M
Mt s ln cc cht c bit c ch mt hoc nhiu bc trong qu trnh sinh tng hp protein
(bng 5.6). Trong s ny c nhng cht c ch chn lc s tng hp protein vi khun m khng nh
hng ti sinh tng hp protein t bo nhn tht. V chng c trin vng s dng nh nhng khng sinh
loi tr s nhim khun m khng nh hng n qu trnh chuyn ho protein ca t bo ch (bng
5.7).
Bng 5.6. Cc cht c ch dch m
Cht c ch Chn lc trn Tc ng giai on
Chloramphenicol T bo nhn nguyn thu Ni di: c ch peptidyl transferase
Cycloheximide T bo nhn tht Ni di: cha r c ch
Erythromycin T bo nhn nguyn thu Ni di: c ch s chuyn peptid ho
Acid fusidic C 2 gii (t bo nhn Ni di: c ch s phng thch ca EF - G

nguyn thu v t bo (hoc EF - 2): phc hp GDP
nhn tht)
Kanamycin C 2 gii Ni di: gy c sai
Neomycin C 2 gii Khi u v ni di: mARN c nhm
Puromycin C 2 gii Ni di: gy kt thc sm
Sparsomycin C 2 gii Ni di: c ch peptidyl transferase
Spectinomycin T bo nhn nguyn thu Ni di: c ch chuyn peptid ho
Streptomycin T bo nhn nguyn thu Ni di: gn vo 30S v nh hng ti tng

tc codon - anticodon gy ra s c nhm ca
mARN
Tetracyclin T bo nhn nguyn thu Ni di: phong b vic gn aminoacyl - tARN

vo v tr A
Bng 5.7. p dng tr liu ca cc cht c ch s tng hp protein
Hp cht p dng v ch dn
Chloramphenicol Ph khng khun rng dng trong iu tr thng hn
Erythromycin Ph khng khun ging penicillin v do thng dng iu tr cc bnh

nhn nhy cm vi penicillin
Acid fusidic Ph khng khun hp, dng cho cc Staphylococci khng penicillin
Kanamycin Dng cho nhim trng gram (- ) nng
Spectinomycin iu tr lao
Nhm tetracyclin Khng sinh c ph khng khun rng, nhng nhiu vi khun khng
vi nhm ny.
V d, chloramphenicol, mt khng sinh c ph khng khun rng. Cht ny v cc cht c ch dch

m c dng nh cc khng sinh, trong khi cc cht khc th ch c dng thm d c ch ca
chnh s dch m.
Tm tt
S dch m l mt qu trnh phc tp vi s tham gia ca nhiu thnh phn:
- ARN thng tin: mang thng tin di truyn m ho di dng codon, mi codon l mt t hp 3
nucleotid m ho cho mt acid amin.
- ARN ribosom: l thnh phn ca ribosom, ni hnh thnh chui polypeptid ang tng hp.
- ARN vn chuyn: yu t mang cc acid amin tng ng ti cc codon trn khun ARN n gn
vo chui polypeptid ang hnh thnh ti ribosom
- Acid amin v ribosom
Qu trnh dch m l cc chu k ribosom, bao gm 3 giai on:
- Khi u: trc tin l hnh thnh mt phc hp gm 3 thnh phn: (1) tiu n v nh ribosom,
(2) tARN mang methionin (Met - tARNiMet), (3) mARN. Mt yu t khi u s pht hin codon khi
u AUG gip phc hp v tiu n v ln ca ribosom gn vo, v s dch m bt u.
- Ni di: cc yu t ni di to ra cc bc chuyn v. Cc acid amin c a n ribosom dng

aminoacyl - tARN theo tng codon ring l.
- Kt thc: khi cc yu t kt thc nhn ra du hiu kt thc trn mARN, s dch m dng li.
S khc bit gia cc thnh phn v c ch chi tit ca s dch m vi khun v t bo nhn tht l
nn tng cho vic s dng rng ri cc cht khng sinh trong vic c ch chn lc qu trnh sinh tng
hp protein vi khun.
CU HI
1. Nng lng gii phng c t GTP thnh GDP + Pi dng :
a) Ghp tARN khi u vi tiu n v 50S
b) Ghp tARN khi u vi tiu n v 30S
c) Dch chuyn ribosome
d) Hot ho acid amin
e) Gn kt mARN vi ribosome
2. Trong qu trnh dch m:
a) Mi tARN c mt tARN - aminoacyl synthetase tng ng
b) Mt tARN - aminoacyl synthetase chung cho tt c acid amin
c) tARN - aminoacyl synthetase ko di chui peptid
d) Mt tARN - aminoacyl synthetase cho mi loi acid amin
e) tARN - aminoacyl synthetase xc tc s chuyn v
3. Trnh t Shine - Dalgarno l:
a) V tr gn ribosom b) Yu t khi lp rp rARN
c) Yu t khi u dch m d) V tr kt thc dch m
e) L trnh t codon khi u
4. Acid amin khi u chui peptid t bo nhn nguyn thu:
a) Formyl - methionin b) Methyl - Methionin
c) Methionin d) AUG
e) GUG
5. Vai tr ca eIF - 2 trong tng hp protein t bo nhn tht:
a) Mang aminoacyl - tARN ti v tr A
b) Gn Met - tARN vo ribosom
c) Hot ho acid amin cn ni di
d) Ti hi EF - Tu
e) Ti hi EF - 1
6. Chui peptid ang hnh thnh gn vo:
a) mARN b) Tiu n v nh
c) Tiu n v ln d) V tr P
e) V tr A
7. S chuyn v ribosom c cc hin tng:
a) tARN vn chuyn xong c tch khi v tr P
b) Peptidyl - tARN di chuyn t v tr A sang v tr P
c) Ribosom tch ra gn vo codon k tip
d) Ribosom chuyn v tng bc
e) a, b v d
8. Yu t no khng tham gia kt thc dch m vi khun?
a) RF - 1 b) RF - 2
c) RRF d) EF - G
e) EF - Ts
9. Tc hi ca streptomycin trong qu trnh dch m ca vi khun:
a) c ch chuyn peptid ho
b) c ch peptidyl transferase
c) Phong b vic gn aminoacyl - tARN vo v tr A
d) Tng tc codon - anticodon gy c nhm ca mARN
e) Gy kt thc sm qu trnh dch m
10. nh hng ca tetracyclin trong dch m vi khun:
a) c ch chuyn peptid ho
b) c ch peptidyl transferase
c) Phong b vic gn aminoacyl - tARN vo v tr A
d) Tng tc codon - anticodon gy c nhm ca mARN
e) Gy kt thc sm qu
Bi 6
IU HO HOT NG GEN
MC TIU
- Tm lc c cc phng thc iu ho hot ng gen t bo nhn nguyn thu.
- M t c c ch iu ho ca t bo vi khun p ng vi cc thay i trong mi trng

sng.
- Gii thch c cch thc tng tc gia cc protein v cc trnh t ADN c bit v cc
phn t khc trong qu trnh iu ho hot ng gen.
6.1. M U
S iu ho hot ng gen vi khun chnh l s iu ho trao i cht, l iu khin tc ca cc

qu trnh sinh ho bng cch thay i thun nghch hm lng cc cht c bn cht protein tham gia vo
cc qu trnh ny hoc l iu ho hot tnh ca chng.
Trong a s cc qu trnh sinh ho, cc cht c dn xut protein u l nhng cht xc tc cc qu

trnh sinh hc, gi l enzym. Song c mt vi qu trnh, v d vn chuyn qua mng sinh hc c thc
hin bi protein, trong protein khng ng vai tr xc tc cho s bin i ho hc m "nhn bit" s
vn chuyn (translocation) c cht.
vi khun c ba loi enzym permease vn chuyn qua mng: permease dipeptid, permease tripeptid
v oligopeptid permease. Oligopeptid permease c oppA, B, C, D, F ln lt chuyn vt cht ln nhau
v qua mng, trong oppA c tnh c hiu.
C hai mc iu ho c bn: mc iu ho sinh tng hp protein v mc iu ho hot tnh

ca chng trong qu trnh thc hin chc nng.
Trong phn ny, chng ta ch xt n s iu ho sinh tng hp protein.
Mc iu ho qu trnh sinh tng hp protein gm nhiu giai on: cc thi k chun b sao chp
b gen v phin m ngha l sinh tng hp cc ARN, thi k kt thc - dch m to ra nhng phn t
protein, mt xch di truyn cui cng ca t bo.
Hnh 6.1. Cc permease vi khun
6.2. IU HO QU TRNH SAO CHP
Qu trnh sao chp, phin m v dch m u gm ba giai on: khi u, ko di v kt thc.
Nhng c im c bn ca qu trnh sao chp ADN l c ch bn bo tn, s gin on (on

Okazaki) v s ko di ca mi ARN trong mi on sao chp, lun lun xy ra theo hng 5 n 3.
Kt thc giai on sao chp, cc on mi (ARNo) b phn hu v cc on Okazaki ni li vi nhau
nh enzym ligase. D nhin, si sm s c tng hp lin tc theo hng 5 n 3.
t bo nhn nguyn thu c h thng a enzym: ADN - polymerase I, II, III, ADN - ligase. Ngoi
ra, li xon v tho xon c helicase v topoisomerase I, phc hi dng siu xon c topoisomerase II
(ADN - gyrase) ph thuc ATP.
Ngoi sao chp b gen, cn c s sa cha v ti t hp, ng thi c s sao chp nhng yu t di
truyn ngoi nhn - plasmid.
Ngi ta xc nh c khong 15 locus di truyn (Dna) kim sot s sao chp ADN. Mi mt gen
nh vy m ho cho mt polypeptid cn thit cho qu trnh sao chp. V d, DnaA cn cho s tng hp
primer (ARNo) v ng vai tr iu ho dng trong qu trnh ny. Song vic to ra sn phm ny li
tn hi n qu trnh t iu chnh. Mt s protein c tnh a chc nng. V d, protein do DnaC tng
hp li cn thit cho vic gn primer vo chui ADN v sau protein ny tr thnh thnh phn ca
phc h sao chp v tham gia vo qu trnh ko di.
Tt nhin s iu ho cht ch hn c xy ra giai on khi ng sao chp. Bn cnh cc protein

ca cc cistron DnaA, DnaC va k trn, cn c s tham gia ca cc protein khc sinh ra t cc locus
DnaB, DnaI v DnaP. S sao chp ADN chu s kim sot dng v kim sot m ca cc protein -
iu ho.
Ni mt cch tng qut, kim sot dng l s tch ly cht hot ho cho n ngng khi
ng mt chu trnh sao chp mi, cn thit cho s cn bng vi vic nhn i ca sinh khi t bo.
Ngc li s kim sot m, khi cht kim hm cn c tng hp mt cch gii hn tip ngay khi
bt u chu trnh sao chp trc (cht km hm nh vy c th l sn phm ca gen nm gn v tr bt
u sao chp, m s phin m ca n ch c thc hin trong giai on sao chp ca vng ADN ny).
Trong qu trnh tng trng ca t bo, cht km hm b yu dn v trong thi im t bo nhn i

th mc km hm ca n h thp di mc ng hn.
6.3. IU HO QU TRNH PHIN M
Trong mi t bo, khng phi gen no cng hot ng nh gen no. Trong khi sn phm ca gen ny
cn c tng hp lin tc, th sn phm sinh tng hp ca gen khc li ch c tng hp trong nhng
giai on cn thit tu thuc mi trng. Thm ch, khi gen "m" th hm lng protein cho chng
cng b kim sot. Do hot ng ca hu ht cc gen u c iu ho trong mt hay mt vi con
ng lm cho hiu ng s dng ca t bo v mt nng lng t c cao nht. C ch iu ho ny
i vi s biu hin gen c th xy ra cc mc khc nhau. S iu ho gen c th xy ra ngay ti
thi im phin m hay trong qu trnh dch m. Sau qu trnh dch m, mt s protein c th b bin i
cu hnh thnh protein chc nng.
Cc gen phin m ARN c gi l cc gen cu trc. Cc protein c dch m t mARN c th l

enzym hay khng phi enzym. Trong s cc protein khng phi enzym c cc protein iu ho. Chng
s gn vi nhng trnh t ADN chuyn bit c gi l operator, kim sot qu trnh phin m ca
cc gen c hiu. Nhng gen tng hp nn cc phn t protein iu ho c gi l cc gen iu

ho. Mi gen cu trc (hay mt nhm gen cu trc) u ng sau mt trnh t (gi l trnh t khi ng)
c nhn bit bi ARN polymerase. Khi polymerase gn vo vng khi ng th s phin m xy ra:
Si c ngha ca ADN s c sao m thnh mARN. Operon l n v phin m bao gm ti thiu l
vng khi ng v cc gen m ho mARN cho mt hay mt vi chui polypeptid. Mt operon c th
cha mt hay nhiu v tr iu ho hn so vi s vng khi ng.
Hot ng phin m ca gen c th khng c iu ho nu sn phm ca chng khng lin quan

ti iu kin mi trng. C th ni nhng sn phm khng ph thuc vo mi trng nh vy c
tng hp lin tc. Tuy nhin s lng ca nhng sn phm "khng c iu ho" ny vn c th thay
i ph thuc vo i lc tng i vi promoter ca ARN polymerase. Cc promoter c i lc cao s to
nhiu sn phm ca gen hn l c i lc thp.
i vi nhng protein i hi mt iu kin mi trng nht nh, th s hot ng ca cc gen to

ra chng thng c iu ho bi mt hay mt s protein iu ho.
Operator l mt trnh t ADN ca operon c mt protein iu ho c gn vo v protein ny

c gi l protein c ch.
Operator c th nm cnh promoter:
Operator c th ph ln mt vng ca promoter: v d operator ca - phage.
S gn ca protein c ch vo operator s ngn cn s phin m ca mt s gen cu trc trong cng

operon. Gen no c dng iu ho ny c gi l gen b iu ho m. Cc operon ca vi khun thng
sn sinh ra cc mARN, gi l a cistron mang m di truyn cho mt s chui polypeptid; th nhng, hu
nh ton b mARN ca t bo nhn tht (tr mt s c quan) u l n cistron.
Cc protein cn thit cho s biu hin ca mt operon c gi l protein hot ho (activator).

Chng c th gn vo v tr khi u nm trong promoter ca operon hay vo trnh t tng cng
(enhancer) nm cch xa operon nhm tng cng kh nng phin m ca cc gen nm trn cng ADN.
Trnh t tng cng u tin tm ra cha 72 bp nm cnh im sao chp ca virus kh 40 (SV 40).
Enhancer c tm thy trong genome ca t bo nhn tht v trong retrovirus. Hot ng ca enhancer
lm tng s lng ARN polymerase khi ADN phin m. Enhancer c th nm cch gen cu trc m
n tng cng.
Hiu qu tng cng c thc hin qua mt loi protein gn vo mt trnh t ADN c hiu. Khi
protein gn c hiu ny gn vo v tr tng cng, cc nuleotid xen gia ni nt ra a v tr tng cng
ln tip xc vi promoter ca operon cn tng cng. Cu trc m nt ny gip cho ARN polymerase
gn c vo gen phin m.
Khi protein hot ho gn vo v tr khi ng hay v tr tng cng s kch thch s phin m ca
gen cu trc trong operon. Ngi ta gi c ch ny s kim sot dng. S kch thch do cc gen iu
ho p ng c th xy ra t nhng phn t nh (ng, acid amin...) n nhng cht ln hn nh cc
phc hp steroid ca hormon (t bo nhn tht) v cc protein th th ca n.
Cht "m" gen, lm cho gen phin m c gi l cht cm ng. Cht "ng" s phin m c gi
l cht c ch. Cc gen c th cm ng thng tham gia trong cc phn ng d ho, v d nh khi phn
gii polysaccharid thnh ng n.
Cc gen c th c ch thng tham gia trong cc phn ng ng ho.
Nh vy c 4 t hp tham gia kim sot s phin m:
- Kim sot cm ng m.
- Kim sot c ch m.
- Kim sot cm ng dng.
- Kim sot c ch dng.
Ta ln lt xem xt tng trng hp.
6.3.1. Kim sot cm ng m
Kiu khi nguyn ca s kim sot m c thc hin bi operon c th cm ng, l "h thng
lactose" ca E. coli.
- galatosidase l mt enzym c hai chc nng. Chc nng ch yu ca n l phn gii lactose
thnh glucose v galactose. Chc nng th hai ca n l bin i lin kt 1 - 4 glycosid ca glucose v
galactose trong ng lactose thnh lin kt 1 - 6 trong ng allolactose. Bnh thng, khi lactose
thiu ht trong mi trng th enzym ny khng nhiu lm. Ngay sau khi thm lactose vo mi trng,
m trong khng c glucose, enzym ny lp tc c sn sinh. Protein vn chuyn c tn l
galactoside permease cn thit cho s vn chuyn c hiu qu lactose qua mng t bo. Protein ny cng
t ngt tng ln sau khi cho lactose vo mi trng. "H lactose" hoang di gm gen iu ho (I+) v
operon cha trnh t promoter (p+), operator locus (O+) v 3 gen cu trc ca - galatosidase (Z+),
permease (y+) v transacetylase (a+). t bin ca tng locus tng ng c tm thy trong t
nhin.
Mt s k hiu:
Cc allen ca promoter
- p+: promoter kiu hoang di, hot ng bnh thng vi ARN polymerase
- p-: promoter t bin khng c nhn bit bi ARN polymerase
- ps: tng cng s nhn bit ca ARN polymerase, nng cao mc phin m ca operon.
- picr: nh hng ti v tr gn ca CRP - cAMP lm gim mc biu hin ca operon lac di 10%
kiu hoang di (icr = insensitive to catabolite repression).
Cc allen ca operator
- O+: allen iu khin ny cho php cc cistron cnh n (tc l v tr cis) biu hin, ngha l c
phin m. Allen ny mn cm vi cht c ch, cht ny ngn chn s phin m phn cn li ca operon.
- Oc: allen iu khin n nh; n khng mn cm vi cht c ch v cho cc cistron cn k phin

m lin tc.
Cc allen - galactosidase
- Z+: m ho mt - galatosidase hot ng khi operon "m".
- Z: t bin c ngha, sinh ra mt sn phm bin tnh khng hot tnh enzym gi l Cz protein.
- Z-ns: t bin v ngha, khng c operon lac no biu hin.
Cc allen permease
- y+: m ho cho mt permease hot ng.
- y: khng m ho permease, c th l t bin v ngha.
Cc allen regulator
- i+: to ra cht c ch hai mt, c th km hm kh nng tng hp ca bt k operon O+ no khi

khng c lactose; khi c lactose th cht c ch b bt hot.
- i-: gen iu ho b hng, khng c kh nng sinh cht c ch hot tnh do t bin v ngha hay
lch ngha.
- is: to cht siu c ch khng nhy cm vi lactose v khng hot ho vi bt k operon O+ no,
ngha l khng b bt hot bi lactose.
i khi c s chng cho gia promoter v cc v tr operator ca "h lac"; trong mt vi operon
ngi ta thy cc locus operator ny ph hon ton ln promoter. Operon iu ho thng xuyn sinh
ra cc protein c ch mc thp, bi v n c mt promoter km hiu sut. Promoter ny c k
hiu bng pi (inefficient promoter). Vic tng hp ra cht c ch khng thay i theo mc lactose trong
t bo. Ngc li, promoter bnh thng (P+) ca operon lactose (lac - p) li gn vi ARN
polymerase rt hiu qu. Khi thiu lactose (iu kin khng cm ng) th protein c ch c hot tnh
(sinh ra bi i+) s gn vo operator O+ (lac O). ARN polymerase lc by gi c th hoc l khng gn
c vo promoter hoc l khng c c operator, bi v protein c ch bao vy c vng promoter.
T lc by gi, s phin m ca 3 gen cu trc ca lac operon b ngn cn.
Khi lactose xut hin (iu kin cm ng c) song li khng c chuyn qua mng t bo vo t
bo cht mt cch hiu qu bi v lc by gi ch mi c vi phn t permease xut hin. Bn trong t
bo, mt vi phn t lactose b bin i thnh allolactose nh - galatosidase. Allolactose l cht cm
ng ca lac operon. N gn vo protein c ch, lm thay i cu hnh ca protein c ch v lm thay i
v tr gn ca protein c ch vo operator. S thay i cu hnh trong phn t protein do mt phn t
khc gn vo, c gi l s bin hnh d lp th. Phc hp allolactose - protein c ch khng th gn
lu vo operator c v n b ri khi ADN ngay (hnh 6.2). Lc ny, ARN polymerase c th c
operator phin m cc gen cu trc trn operon. Khi lng permease tng ln, lactose c chuyn
qua mng nhiu ln v ng lactose li b phn hu bi - galatosidase. Khi lactose trong mi trng
tr nn cn kit, protein c ch va c tng hp s c gii phng khi allolactose, do chng
(cht c ch) c th gn cht vo operator, lm ngng phin m ca cc gen cu trc trong operon lac.
Cht c ch ni chung l cc protein c cu trc oligomer, trong mi mt tiu n v ca n u c
hai tm c th. Trong hai tm ni trn, tm no c i lc cao vi trnh t ca ton b promoter th c
coi l tm gn cht c ch, cn tm cn li s l tm gn cht cm ng. S gn cht cm ng vo cht c
ch tm bin cu th hai s lm thay i cu hnh ca cht c ch, chnh v vy m lm gim i lc
ca tm gn cht c ch i vi operator v gip operator t gii thot khi cht c ch.
Allolactose lun c xu hng tch khi protein c ch, ngay sau khi mc lactose trong t bo xung
thp. Thm ch khi "h lac" b c ch, i khi protein c ch tch khi operator mt cch nhanh chng.
ARN polymerase c th ln vo operator m operon v tng hp nn phn t mARN a cistron. iu
ny gii thch c mc rt thp ca permease v - galatosidase lun lun thng trc trong t bo
vi khun.
Cc phn t mARN vi khun c thi gian bn hu rt ngn (ch mt vi pht), do s tng hp

protein ngng li rt nhanh sau khi phin m b c ch. Mt khc, protein rt bn vng, nhng li d
phn tn mi khi t bo phn chia. Operon cha gen iu ho (i) trong h lactose bao gm promoter - i
v gen cu trc ca protein c ch. Bnh thng promoter ny c hiu lc rt km v ch c mt t phn
t protein c ch - lac tn ti trong t bo. Tuy nhin, cc operon nm st vi promoter ca gen iu
ho v s t iu ho c th xy ra.
Hnh 6.2. S cc yu t chnh kim sot lac operon
Hnh 6.3. Qu trnh cm ng enzym
(a) protein c ch gn vo operator v phong b s hot ng ca ARN polymerase;

(b) cht cm ng gn vo protein c ch v bt hot n. ARN polymerase tin hnh phin m cc
gen 1,2,3 trong operon
6.3.2. Kim sot c ch m
Ngoi c ch cm ng m (hoc gii c ch) va nu trn, cn c s thch ng enzym m hay cn

gi l c ch m qu trnh tng hp enzym. Trong trng hp ny, s tng hp enzym thay v c gy
cm ng bi c cht, th n b c ch bi chnh bn thn sn phm m n xc tc. Hin tng ny c
quan st thy trong trng hp tng hp acid amin tryptophan v histidin. Tryptophan c tng hp
theo 5 bc, mi bc c xc tc bi mt enzym c hiu (hnh 6.4). Cc gen p ng cho 5 enzym
nm trn mt operon, thng thng theo mt trt t ging nh trt t ca sn phm enzym ca chng
trong qu trnh sinh tng hp.
Gen iu ho cho h thng ny lun lun sn sinh ra mt loi protein khng chc nng, c gi l
cht c ch gc. Khi tryptophan sn xut d tha, th s qu ti ca tryptophan s tc ng nh mt cht
ng c ch. Cht ng c ch gn vo cht c ch gc hnh thnh nn mt phc hp c ch chc nng.
Phc hp c ch chc nng ny gn vo operator trp, v c ch s phin m ca 5 gen cu trc trn
operon. Cc vng promoter v operator ca operon ny nhiu khi ph ln nhau v gn c phc hp co
- aporepressor, do ARN polymerase kh chen vo c. Khi nng tryptophan gim xung,
tryptophan tch khi cht c ch gc v cht c ch gc ri khi operator, th ARN polymerase li tng
hp c mARN a cistron cho 5 enzym tng hp tryptophan.
Hnh 6.4. Kim sot c ch m trong h tryptophan ca E.coli
6.3.3. C ch iu ho suy gim
C ch iu ho th hai ny c pht hin h tryptophan. Ti u 3 ca operon phin m mARN

a cistron tng ng g1, g2, g3, g4, g5 c mt trnh t gm 162 base, phn b gia operator v gen cu
trc th nht, c gi l trnh t dn (hnh 6.5, hnh 6.6).
Hnh 6.5. S phn b gen ca tryptophan operon
Hnh 6.6. Sp xp cc vng nucleotid trong trnh t dn
Trnh t dn cha mt phn ng vai tr b suy gim, chnh b suy gim ny m ho phn t peptid
dn cha 14 acid amin bo hiu cho s phin m dng li.
Trnh t dn cha 4 vng, trong vng A cha hai codon m ho cho 2 codon m sao cho
tryptophan (UGGUGG) c k hiu XX. Phn D trc khi ni vi Tryptophan operon c mt trnh t
m ho cho 7 - uracyl trn mARN.
ARN polymerase chuyn ng dc si c ngha ADN theo chiu 3 5 v phn t mARN tng
ng s c tng hp theo chiu 5 3. Gn u 5 ca mARN l v tr gn ribosom (hnh 6.7).
Hnh 6.7. S chuyn dch ca ribosom trn mARN
Ribosom gn nh chuyn dch lin tip theo sau ARN polymerase trong qu trnh phin m v gii
m. Ribosom gn v trt trn mARN. Cc vng A v B c th bt cp vi nhau v c cha nhng trnh
t b sung v vng C, D cng vy. Nhng nu B cng bt cp c v tr vi C th s c th hnh thnh
nn 3 nt kp tc A/B, B/C v C/D.
Trong iu kin tha tryptophan th ribosom dch m ra si peptid dn, do vng B v C ca

mARN khng to nt c. Nh , vng C v D bt cp c vi nhau to thnh nt kp tc ngng
phong b ARN polymerase. Khi Tryptophan operon khng hot ng. Nu mi trng thiu
tryptophan th ribosom s dng li cc codon tryptophan ca vng A.
V ribosom ph ln A, nn B bt cp vi C, do vy C khng th bt cp vi D nn khng hnh thnh

c kp tc ngng. Nh , ARN polymerase tip tc phin m cc gen cu trc ca tryptophan
operon to nn mARN tng hp cc enzym cho tryptophan (hnh 6.8).
S phin m suy gim ca tryptophan operon cng nhy cm vi cc nng ca mt s acid amin
khc ngoi tryptophan nh histidin v leucin.
Hnh 6.8. M hnh phin m suy gim ca operon tryptophan E. coli

(a)Tha tryptophan: ngng; (b)Thiu tryptophan: khng ngng
6.3.4. Kim sot cm ng dng
a s cc h thng tng hp enzym cm ng c bit cho n nay u hot ng nh s iu ho

m tnh, ging nh nu trong v d d ho lactose. Khc vi s iu ho tng hp enzym m tnh
trong qu trnh trao i lactose, cc enzym ca qu trnh trao i arabinose E. coli nh s kim sot
ca repressor c kh nng thc hin iu ho dng tnh v m tnh. Arabinose l ng m s chuyn
ho ca n i hi 3 enzym c m ho bi cc gen araB, araA, araD. Khi mi trng khng c
arabinose th 3 enzym trn ch c mt lng rt nh. Khi b sung arabinose vo mi trng nui cy
hm lng 3 enzym ny cng tng ln. Qu trnh cm ng c iu ho bi mt gen araC nm gn 3
gen trn nhng cch chng bi phc hp iu ho (gm operator (araO) v initiator (araI), araI cha
promoter ca nhm BAD.
Sn phm protein ca araC (AraC) l cht c ch ca nhm BAD khi mi trng khng c
arabinose. N lun lun gn vo araO lm ADN c dng loop nn ARN polymerase khng gn vo
c phin m. Tuy nhin, sau khi gn vi arabinose th repressor ny b bin i cu hnh thnh
phc hp hot ho ri khi operator (araO) v gn vo initiator (araI), to iu kin cho ARN
polymerase gn c vo promoter ca BAD, nh cm ng c s phin m ca operon. Bn thn
AraC t iu ho chnh n bng cch gn vo araC.
Cn lu rng, operon ca arabinose c coi l mt trong s cc operon nhy cm i vi glucose

v do s tng hp mARN ca n cng ph thuc vo lin kt ca CAP l mt protein hot ho gen d
ho vi promoter BAD. Ni cch khc cho operon ca arabinose hot ng mnh cn c hai tn hiu
iu ho dng tnh, tn hiu th nht thng bo s c mt ca arabinose, tn hiu th hai s vng mt
ca glucose.
Hnh 6.9. Kim sot cm ng dng operon arabinose
Adenosine monophosphat vng (cAMP), protein hot ho gen d ho (CAP) v protein nhn AMP
vng (cyclic AMP receptor protein = CRP) tham gia vo s iu ho ca h thng arabinose. AMP vng
c kh nng hot ho cho nhng promoter c i lc yu nhng bn thn n khng gn c vo
promoter. Mun lm iu ny n phi lin kt vi CAP hoc vi CRP. Khi AMP gn c vo promoter
n s hot ho ARN polymerase v ARN polymerase s gn c vo promoter.
6.3.5. a kim sot
Mt locus di truyn c th c kim sot bi nhiu c ch khc nhau. Khi c mt glucose th vi

khun E. coli khng cn thit phi phn gii mt loi ng no khc v cc gen m ho cho cc enzym
phn gii cc ng y cng ng li. V d, nu khng c glucose nhng c lactose trong mi trng
th lac operon c cm ng. Nhng nu c glucose th s cm ng ca lac operon s khng xy ra.
Hin tng ny c t tn l hiu ng glucose (glucose effect), ngy nay ngi ta gi hin tng ny
l c ch d ho (catabolite repression). Phc h ca 2 cht hot ho c ch d ho l cAMP v CAP.
Vi lac operon (hnh 6.2) y c v tr gn phc h cAMP - CAP. ARN polymerase ch gn c

hiu qu vo promoter nu phc hp cAMP - CAP cng c gn vo v tr ny. Khi mc glucose tng
ln trong t bo, th lng cAMP gim xung v s c t phc h cAMP - CAP hn c th dng hot
ho lac operon. Bn thn cAMP khng gn c vo promoter ARN polymerase d gn vo ADN,
n phi lin kt vi CAP lm thay i cu hnh ca CAP. Lc phc h cAMP - CAP gn c vo
promoter. S iu ho ny l s iu ho cm ng dng tnh.
CAP c sinh ra mc thp do mt locus di truyn c hiu. Enzym adenylate cyclase

(adenylcyclase) bin adenosine triphosphat (ATP) thnh adenosine monophosphat vng (cAMP).
6.4. KIM SOT SAU DCH M
S biu hin ca gen c th c iu ho sau khi protein hnh thnh.
S km hm ngc cn gi l s c ch bi sn phm cui, cng l c ch iu ho c s tham gia

c ch ca enzym. Sn phm cui cng ca qu trnh sinh tng hp (thng l phn t nh nh acid
amin chng hn) c th lin kt cht ch (nu nng cao) vi enzym u tin c tng hp ra. S
lin kt ny khng xy ra ti v tr xc tc (trung tm hot ng) ca enzym m v tr gn cht km
hm (v tr iu ho) lm bin tnh cu trc bc 3 hay bc 4 ca enzym, do bt hot v tr xc tc.
S bin hnh d lp th ca enzym s phong b kh nng xc tc ca enzym v ngn cn s sn xut

qu ca sn phm cui cng trong qu trnh sinh tng hp cc cht chuyn ho trung gian.
Hnh 6.10. S km hm ngc trong E. coli

(e = enzym; g = gen; 1,2,3,4,5 = cc cht trung gian)
V d, sn phm cui cng isoleucin trong E. coli khi xut hin nng cao, lin kt vi enzym
u tin (e1) ca qu trnh sinh tng hp v do vy km hm ton b con ng ny cho n khi
isoleucin tr li mc bnh thng thng qua mc tiu th ca t bo.
Cht khi u ca chui gm 5 phn ng ny l threonin. Phn ng u tin threonin bin thnh
acid - ketobutyric do enzym threonin - desaminase, qua mt lot qu trnh chuyn ho to ra sn phm
cui l isoleucin. Vo mt thi im no , isoleucine khng c t bo s dng n s lin kt tr li
vi threonin-desaminase v km hm tc dng enzym ny. Cc enzym tuy vn c hnh thnh

nhng khng hot ng c. Vic tng hp isoleucin b nh ch cho n khi t bo dng ht acid amin
tha.
Tm tt
Vi khun sng trong mt mi trng cnh tranh gay gt, chng cn phi thay i nhanh chng trong
c ch chuyn ho sng st. Nhiu vi khun to ra cc phng thc chuyn ho thch ng vi s
thay i ny, cc enzym trong qu trnh chuyn ho ch c tng hp khi cn thit. S kim sot nh
vy c thc hin mc gen.
Cc enzym cn cho cc phng thc d ho ch c to ra khi c sn mt cht dinh dng c bit

nh lactose. Cc protein c ch km hm s biu hin gen v tc ng c ch b km hm bi cc cht
cm ng, cc cht cm ng bn thn n l cc phn t dinh dng hay cc phn t c ngun gc t cht
dinh dng. Cc kim sot ny b thay i nu c s la chn gia glucose hay lactose th glucose c
u tin hn.
phng thc ng ho, cc enzym cn cho s sn xut mt acid amin c bit nh tryptophan ch
c tng hp khi tryptophan khng c sn vi khun. trong trng hp ny, cc cht c ch l sn
phm cui cng ca qu trnh.
Cc phng thc khc ca s kim sot biu hin gen l bng cc protein hot ho, y l s kim
sot dng. C s tham gia ca cAMP vo vic iu chnh s biu hin gen vi khun.
CU HI
1. Protein c ch khc vi protein hot ho ch:
a) Gn vo operator ngn cn phin m gen cu trc
b) Thuc dng iu ho m
c) Gn vo v tr khi u trn promoter
d) Gn vo v tr tng cng
e) a, b
2. Hai chc nng ch yu ca enzym - galactosidase th hin :
a) Phn gii lactose thnh glucose v galactose
b) Phn gii lactose thnh glucose v fructose
c) Bin i lin kt 1 - 4 glycosid ca glucose v galactose thnh 1 - 6 trong allolactose
d) Bin i lin kt 1 - 6 glycosid trong allolactose thnh lin kt 1 - 4 trong lactose
e) a, c
3. "H lactose" hoang di ca E. coli c:
a) Gen iu ho (Is)
b) Operon cha promoter (Picr) v operator (Oc)
c) Operon gm vng promoter v enhancer
d) 3 gen cu trc: - galactosidase (Z+), permease (y+), transacetylase (a+)
e) a, b, d
4. "H lactose" thng xuyn sn sinh protein c ch mc thp v:
a) E. coli chung lactose hn glucose
b) Promoter ca operon ny km hiu sut
c) Promoter ca operon ny gn vi ARN - polymerase
d) Cht c ch c tng hp trong t bo c thay i
e) a, b ng
5. Cht c ch gc l:
a) Protein khng chc nng sinh ra do gen iu ho ca h tryptophan
b) Tryptophan
c) 5 enzym tng hp tryptophan
d) Trnh t dn
e) 5 cht chuyn ho do 5 enzym tham gia tng hp tryptophan
6. Operon ca arabinose c coi l operon nhy cm i vi glucose v
a) AMP tng khi hm lng glucose tng
b) AMP vng c kh nng hot ho promoter yu
c) AMP mun gn vo promoter phi nh CAP
d) AMP gn c vo promoter s hot ho ARN polymerase
e) b, c v d
7. S km hm ngc khng chp nhn trng hp:
a) S c ch bi sn phm cui cng
b) C ch iu ho c s tham gia c ch ca enzym
c) S lin kt gia sn phm cui cng vi enzym v tr iu ho ca enzym lm bt hot v tr

xc tc
d) S lin kt gia sn phm cui cng vi enzym v tr xc tc ca enzym
e) S bin hnh d lp th ca enzym s phong b kh nng xc tc ca enzym
8. Operon gm:
a) Vng khi ng (promoter)
b) Cc gen cu trc
c) V tr iu ho
d) Chp GMP
e) a, b v c
9. Operator l:
a) on mARN gn c protein iu ho
b) on ADN chuyn bit gn c protein iu ho
c) on ADN nm trc promoter
d) on ADN nm sau promoter
e) Gen tng hp protein
10. Kim sot dng khc vi kim sot m v cn phi:
a) Loi b tch cc phn t c ch
b) Hot ho qu trnh khi u ca ARN - polymerase
c) a vo co - repressor
d) Loi b co - repressor
e) a v d
Bi 7
B GEN T BO NHN THT
MC TIU
- So snh c b gen ca t bo nhn tht v t bo nhn nguyn thu.
- M t c cc loi trnh t chui khc nhau tm thy b gen t bo nhn tht.
- Nu c s biu hin ca b gen t bo nhn tht.
- Nu c cc cch kim sot s biu hin ca b gen.
7.1. M U
Cc b gen t bo nhn tht phc tp hn t bo nhn nguyn thu. S gen m ho cho protein cng
nhiu hn gp trm ln. S d ADN c th hin nhiu loi trnh t khc nhau. ADN ca t bo nhn
tht dng thng, cn t bo nhn nguyn thu th dng vng.
im khc na l s phin m v dch m t bo nhn tht c phn ra lm 2 giai on tm thi

v mt khng gian: s phin m xy ra trong nhn, cn s dch m xy ra trong t bo cht. Ngc li
vi t bo nhn nguyn thu: s dch m ca phn t mARN c th bt u trc khi mARN c tng
hp xong.
7.2. T CHC B GEN T BO NHN THT
7.2.1. Kch thc ca b gen t bo nhn tht - gi tr C
Gi tr C (C - value) c dng din t kch thc b gen ca mt loi, n biu th s cp

nucleotid (thng bin din bng n v Mb hay Mbp - megabase pair) ca b gen n bi ca mt sinh
vt t bo nhn tht. Cc t bo t bo nhn tht cha nhiu ADN hn l cc t bo nhn nguyn thu
(bng 7.1).
Bng 7.1. B gen cha acid nucleic
B gen S lng gen S cp base
Thc vt <50 000 <1011
Ngi 25 000 3.109
Giun 14 000 108
Rui 12 000 1,6.108
Nm 6 000 1,3.107
Vi khun 2 000 - 4 000 <107
Mycoplasma 500 <106
ssADN virus
Parovirus 5 5 000
Phage fx174 11 5 387
dsARN virus
Coronavirus 7 20 000
Infuluenza 12 13 000
TMV 4 6 400
Phage MS2 1 1 300
Viroid
PSTV ARN 0 359
Nghch l gi tr C l mt khi nim phn nh s khng tng xng gia kch thc b gen (gi tr
C) vi s gen ca mt loi. V d, ngi c b gen ln hn giun n 30 ln nhng ch gp i v s gen.
Nh vy phn ln trnh t tn ti trong b gen khng lin quan n gen.
7.2.2. S khi qut v cc loi trnh t ADN
7.2.2.1. S khng ng nht ca ADN t bo nhn tht
Khi tch ADN ca t bo nhn tht bng siu li tm trn thang nng Cesium chlorid, c 3 vt
c ghi nhn (hnh 7.1).
Hnh 7.1. S xut hin ca ADN v tinh sau khi siu ly tm
S khng ng nht ca ADN t bo nhn tht th hin r hn khi thc hin phn ng ti hp
(reassociation). ADN c ct nh v cho bin tnh ri sau cho hi tnh. Khi hi tnh, cc on c
trnh t b sung d ti hp vi nhau, nh vy nhn bit cc trnh t lp li.
ng hc ti hp thnh cp ca ADN t bo nhn tht khc vi t bo nhn nguyn thu (hnh 7.2).
ng cong ti hi ca t bo nhn nguyn thu c dng hnh sigma in hnh, chng t s ng nht
ca cc trnh t trong ti hp. t bo nhn tht ng cong phc tp hn, ko di mt khong rng cc
gi tr C0t (n v ca n l s mol nucleotid/lit/giy) v c cha 3 thnh phn lp li mc khc
nhau:
- ADN lp li nhiu (ti hp rt nhanh), chim khong 10 - 15% b gen.
- ADN lp li trung bnh (ti hp nhanh va), chim khong 25 - 40% b gen.
- ADN duy nht (ti hp rt chm), chim khong 60% b gen.
Phc hp ca mt vi b gen bit kch thc c th c tnh ton bng cch so snh ng cong
C0t ca n vi ng cong C0t ca mt ADN mt phc hp c bit trc (thng l ADN E. coli
vi kch thc b gen c tng chiu di ca cc trnh t l 4,2.106 bp). Cch so snh nh vy s dng
ca mt dn s ADN, l gi tr C0t thi im 50% ADN ti t hp.
C0t 1/2 ca ADN t b gen th nghim = phc hp hin din ca b gen th nghim.
C0t 1/2 ca ADN E. coli = 4,2.106 bp.
Hnh 7.2. ng cong ti hp ca ADN ng vt c v
1. ADN c kh nng bt cp tc thi; 2. ADN bt cp chm hn 10 - 3<C t<10;

0
3. ADN bt cp rt chm 10 < C0t < 103
7.2.2.2. Cc trnh t ADN lp li mc cao
Cc trnh t ny cng c bit nh l ADN v tinh (satellite DNA), bi v khi ADN ca b gen
c ct ra v ly tm trong cc nng gradient t trng cesium chlorid. Cc chui lp li cao thng
hnh thnh cc di bng v tinh cch xa nh chnh (kh rng) so vi cc chui lp li trung bnh hay
duy nht. iu ny cho thy rng ADN lp li cao thng c mt thnh phn base khc vi thnh phn
ca b gen (v v vy t trng cc phn ni khc nhau).
Cc trnh t ny chim 10 - 15% b gen ca ng vt c v v khng m ho cho protein. Chng

lin quan n cc cht d nhim sc c cu (constitutive heterochromatin).
Phn tch trnh t nucleotid cho thy chng gm 3 loi:
- H ADN vi v tinh: gm cc on nh ca trnh t lp li lin tc (tandem) c trnh t n gin

(thng 1 - 4 bp) v nm ri rc khp b gen. Trong trng hp lp li 1 nucleotid, A v T thng gp,
chim khong 10 Mb hay 0,3% b gen t bo nhn tht. Tri li, G v C rt him gp. Trong trng hp
lp li 2 nucleotid, on lp li CA (GT trn si b sung) thng gp, chim khong 6% b gen v
thng a hnh. on lp li CT/GA cng ph bin, thng xy ra trn mi 50 kb v chim khong 2%
b gen, nhng CG/GC rt him. iu ny do C b sung vi G u 3 b methyl ho, kt qu to TpG
(hay CpA ca si b sung). on lp li 3 v 4 nucleotid kh him, thng a hnh v c nghin cu
dng lm cht nh du a hnh. im ni bt ca ADN vi v tinh cha c bit. Cc on lp li xen
k purin - pyrimidin, nh cc on lp li lin tc ca cp 2 nucleotid CA/GT, c kh nng ph hp cu
to ca ADN xen k, Z - ADN, in vitro, nhng t c bng chng chng cng lm nh vy trong t bo.
- Loi th hai: tng ng vi cc trnh t lp li tandem nhng vi cc on di hn (100 - 200 bp).
- Loi th ba: c nhiu trnh t lp li mc cao, phn tn pha ngoi cht d nhim sc nh trnh
t CEN v trnh t TEL.
+ Trnh t CEN: cc trnh t CEN lp li cao l ca cc tm ng (centromere). nm men

Saccharomyces cerevisiae gm:
Trnh t u c 9 cp base 5TCACATGAT.
Trnh t gia c 80 90 cp base, rt giu A v T (>90%).
Trnh t 11 cp base u 3TGATTTCCGAA.
Cc trnh t ny ADN ngi phc tp hn.
- Cc trnh t TEL:
Cc trnh t TEL thuc cc nhm telomere (u mt ca nhim sc th) vi nhiu vai tr khc nhau
nh bo v u mt nhim sc th khi b ct, gi chiu di ca nhim sc th khi sao chp, gn vi
mng nhn v km hm s biu hin ca cc gen u mt.
Cc trnh t TEL c tnh bo tn trong tin ho. Chng c s ln lp li cao, giu C v A: CC(C)
ACACA(CA) nm men, CCCTAA ngi. u mt ca nhim sc th giu G gp li hnh kp tc c
cu trc 4 mch. Cu trc ny bo v u mt nhim sc th khi b ct bi nuclease, ng thi khi sao
chp gi u mt khi b mt cc trnh t m ho.
Cc chui v tinh lp li cao thng thy trong tt c cc nhim sc th ca b gen. V d, 25% b

gen ca Drosophila virilis c trnh t ACAAACT, hin din khng t hn trong 107 bn sao.
7.2.2.3. Cc trnh t lp li mc trung bnh
Loi trnh t ny chim 25 40% b gen ngi. Chng cng gm cc trnh t lp li nhng di hn
(100 1000 bp) v a dng hn nhiu so vi loi lp li cao (v tinh). Cc trnh t ny phn tn trong b
gen v nu ct b gen thnh on 20 40 kb th c 90 n 100% s on c trnh t lp li trung bnh.
Trong s cc trnh t lp li trung bnh, c cc h trnh t c hiu l SINE v trnh t LINE.
Mt t bo nhn tht c th c hng trm nhm ADN lp li trung bnh, vi mi nhm c khong t
50 - 105 on lp li.
C mt s lng ln cc nhm khc ADN lp li trung bnh trong b gen ngi, nh chui 6400 cp
base c lp li t 3000 - 4600 ln c tnh cho khong 1% trong tng s ADN ngi.
- Cc trnh t SINE (cn gi l Alu ngi)
Mt trong nhng nhm ln nht ca ADN lp li trung bnh ngi l phc hp ca cc chui c
gi l h Alu. S d c tn ny v n c cha v tr nhn din cho enzym ct gii hn AluI. Chui Alu
xut hin khong 3x105 ln trong b gen n bi ca ngi, v vy chim khong 3% ADN ngi.
Alu cha mt lng ln GC v mc d nm ri rc khp cht nhim sc hot ng (euchromatin)

ca b gen, nhng chng thng trn nhim sc th R v thy trn di xanh khi dng thuc nhum
Giemsa chun v cha cc vng c kh nng phin m ca b gen. Chiu di ton b ca on lp li
Alu khong 280 bp v thng tip theo bi cc trnh t thun (6 - 18 bp) (c cng hng), giu A ti
mt si v T ti si b sung. Tuy nhin, c s khng i xng ti cc on lp li khng lin tc: mt
n v lp li cha trnh t 32 bp bn trong nhng on khc khng c. Cc monomer cha mt trong
hai on lp li lin tc v kiu gen khc nhau ca dimer v monomer.
S chuyn v nghch to bn sao ca cc phn t v tr mi, trong khi phn t cho ban u vn gi
nguyn cu trc khng bin i. Do vy, chuyn v nghch to nn mt t on t v s ti cu trc ca
b gen t bo ch. Nhng bin i ny s lm ngng hay hot ho cc gen, m mt s bin i ny c
th gy ra ung th.
Hai n v lp li ca trnh t Alu ging nhau v trnh t 7SL ARN, thnh phn cu to ca du hiu
nhn din, vn chuyn protein qua mng li ni sinh cht. Do iu ny ngi ta tin rng trnh t Alu
tng ln l nh s chuyn v 7SL ARN, do vy to thnh gen gi ct 7SL ARN. Qu trnh chuyn v
trnh t Alu xy ra nh enzym phin m ngc m ho bi LINE - 1 (Kpn) v c th l nguyn nhn gy
cc vn lm sng. C th s lng ln cc bn sao gen gi ct lin quan n s hin din trnh t
promoter trong trnh t 7SL ARN (gen 7SL ARN, ging gen tARN c m ho bi ARN polymerase
II t mt promoter ni). Ngc li, gen gi ct t bn sao ARN polymerase II thiu trnh t promoter v
chng ch c th biu hin nu chng gn trnh t promoter c chc nng.
Hin nay chc nng ca trnh t Alu cha uc bit. Mc d tn s thng gp l 1 bn sao trn 4
kb, nhm cc on lp li Alu xy ra ti cc vng bt k. Do chng c mt khp ni, trnh t Alu

c xem nh thc y qu trnh ti t hp khng tng ng, nguyn nhn gy bnh trong mt s
trng hp nhng cng c th l cc tin ho do thc y nhn i ADN. Mc d thng thiu mt trnh
t m ho, trnh t Alu thng tm thy ti cc v tr khng m ho ni sinh, ti intron v cc trnh t
khng dch m. Do vy, chng thng hin din trong bn phin m ARN u tin t gen m ho
polypeptid, thng l mARN. In vitro, trnh t Alu cng c th c sao chp t promoter ni nh ARN
polymerase III v bn sao ca mt vi trnh t Alu do s tch ly cc ARN t bo cht c th lin kt vi
protein ca du hiu nhn din.
Hnh 7.3. Cc trnh t SINE
- Cc trnh t LINE
Chng gm cc h LINE 1 hay KpnI v THE 1, xy ra mi 50 kb trn b gen ngi. Cc trnh t

LINE c to ra do ct b gen vi enzym gii hn KpnI. Chng c chiu di khong 6 v 7 Kb vi gn
5 000 bn sao nguyn vn v 100000 bn sao tng phn ri khp b gen ngi. Chng l nhng trnh t
lp li khng m ho di nht v thng vng giu AT, ging nh cc trnh t Alu (hnh 7.3). Cc bn
sao m trnh t LINE gn vi protein to thnh phc hp ribonucleoprotein. mt dng t bo ngi b
ung th vng mc (teratocarcinome), ngi ta quan st thy cc ribonucleoprotein ny. S xen on
LINE vo cc v tr khc nhau c th gy hu qu nht nh, nh trong trng hp bnh mu khng
ng A (hemophilia A).
Trong h LINE 1, nhiu on c kh nng chuyn v. Chiu di ton b l 6.1 kb v c 2 ORF, tuy
nhin chng khng hin din trong hu ht trnh t ring r. ORF 1 gn cui (u tn cng 5) v m
ho protein cha r chc nng p40 (trng lng phn t khong 40 kDa). ORF2 c nhng vng ging
vi trnh t nucleotid m ho cc enzym phin m ngc khc nhau v cc protein virus khc. Cu trc
hon chnh ca LINE 1 cha mt promoter trong vng ADN khng dch m trc ORF1 (gi l 5 -
UTR) trong khi ti u 3 c trnh t (A)n/(T)n, thng c ui poly A. Trong trng hp cc cu trc
khc c th chuyn v v cc cu trc LINE - 1 b kp bi cc trnh t i ngn. Cu trc LINE - 1 hon
chnh kh him (ch khong 3500 bn sao) v hu ht cc on lp li b ct u 5 do vy chng c
chiu di khc nhau v thng c ui polyA. Cc cu trc LINE - 1 thng vng cht nhim sc
nhng khc vi trnh t lp li Alu do thch nhnh G ti (Giemsa dng) ca nhim sc th k gia.
Ging Alu, chng thiu cc trnh t m ho nhng c th tm thy cc trnh t khng m ho ni sinh.
Do vy, chng hin din trong bn sao ARN u tin ca cc gen ln nhng khng c trong mARN.
Hnh 7.4. Trnh t LINE
- Cc gen ca rARN, tARN, ARN 5S v ARN 7SL
Mt lot cc gen gi vai tr quan trng trong sinh tng hp protein nh cc gen ribosome, tARN,
ARN 5S v ARN 7SL c s lp li hng nghn ln.
+ Gen nhm I: cc gen ca rARN
Cc gen rARN c phin m bi ARN polymerase I. Bn phin m u tin l pre - rARN, sau khi
ct ni to ra 3 loi rARN l rARN 28S, rARN 18S v rARN 5,8S. Cc gen ny khng phn tn m xp
thnh cm (cluster), mi cm c th hn 200 bn sao. ngi, cc cm c tm thy trn vai ngn
ca cc nhim sc th tm u (acrocentric) 13, 14, 15, 21 v 22, chim khong 0,4% b gen. Cc nhm
ny xp quanh yu t t chc hch nhn hnh thnh kiu cu trc c bit trn nhim sc th v gian
k to nn cc hch nhn (nucleolus).
Cc gen 28S, 5,8S v 18S rARN bt thng do cc gen nhn chim s lng ln, c phin m
ring r, u tin chng c biu hin nh l cc bn sao a gen, theo cch ca gen ty th. on sao
chp ca 13 kb biu hin tin 45S rARN sau tri qua nhiu phn ng khc nhau to thnh 28S,
5,8S v 18S rARN trng thnh. Ging nh cc sn phm ca gen ty th, cc sn phm rARN ring r
ca nhm rADN l cc thnh phn c lin quan vi mt chc nng ring bit. Do vy, vic s dng bt
thng cc bn sao ARN khng khc so vi cc sn phm dch m u tin t t c tch ra thnh 2
hay nhiu chui polypeptid c chc nng thng thng. V d, phn t insulin c 2 chui polypeptid
c to thnh t sn phm dch m duy nht. Cch di truyn ny c lin quan n cc sn phm khng
trng lp t tin phn t duy nht him khi xy ra: cc thnh phn ring r ca a s protein a tiu n
v ngi c m ho bi cc gen khc nhau, thng trn cc nhim sc th khc nhau.
+ Gen nhm III: cc gen tARN, ARN 5S v ARN 7SL
Cc gen ny c phin m bi ARN polymerase III nn gi l nhm III. Nhm ny gm cc gen

m ho cho mt s ARN nh tm thy trong nhn v t bo cht.
ngi c hn 200 gen m ho cho ARN 5S. Cc gen ny tp trung thnh cm mt s im nht
nh ca cht nhim sc th. mt s loi nh Xenopus s bn sao ca ARN 5S c th hn 20 000.
7.2.2.4. Cc trnh t c nht
Cc trnh t c nht chim phn ln b gen (chim khong 60%), gm 2 loi:
- Cc gen m ho cho protein.
- Cc gen gi.
Cc gen m ho cho protein thuc nhm II v n c phin m do ARN polymerase II. Tr cc gen
m ho cho protein histone, cc gen nhm II c trnh t c nht hoc vi s t bn sao. Cc gen nhm II
ny hu nh ch m ho cho mt loi protein.
a s chng c trnh t c nht hay gn nh c nht. Mt s gen c bn sao th hai trong qu

trnh tin ho. C hai bn sao ny c th chuyn i b tr nhau, nh trng hp cc gen ca globine.
i khi, hai bn sao c s phn ho nh trong tin ho ra hai protein nhng t khc nhau.
Mt gen c th to ra c nhiu bn sao rt sm trong qu trnh tin ho. Mi bn sao c s phn

ho c lp nhau do hin tng nhn i (duplication). S phn ho ny dn n hng lot gen m ho
cho cc protein tng ng. S biu hin ca cc gen ny ph thuc vo kiu hay trng thi ca t bo.
Cc gen nh vy hin din dng lp li thnh h. V d, nhiu h gen c bit nh:
- H cc gen Globine.
- H cc gen Actine.
- H cc gen Myosine.
Cc gen Globine tp hp thnh cm: phc hp ca cm Globine nm trn vai ngn ca nhim sc
th 16 v phc hp Globine nm trn vai ngn ca nhim sc th 11. Trnh t sp xp ca gen tng
ng vi trnh t biu hin trong qu trnh pht trin c th.
Hemoglobine l protein c cu trc bc bn. ng vt c v trng thnh, cu trc bc bn gm 2

mch polypeptide v 2 mch .
Trong qu trnh pht trin c th c s thay i cc dng cu phn nh bng 7.2.
Bng 7.2. Mch Globine ngi trong qu trnh pht trin c th
Peptide globine Phi Thai Ngi ln
- tng t (xi)
- tng t (epsilon) G , A ,
Gen gi c xc nh bi vic cha cc trnh t c lin quan mt thit vi cc gen cu trc, nhng
khng dch m thnh protein chc nng c.
S sp xp ca cm cc gen - tng t globine ca th c trnh by hnh 7.5. C 4 chui -

tng t globine trong nhm ny. Gen 1 c biu hin cc th trng thnh, trong khi cc gen 3
v 4 th c hot tnh ch yu trong qu trnh pht trin ca phi.
Phn tch trnh t 2 cho thy rng mc d gen 2 c quan h mt thit vi cc trnh t khc trong
cm, nhng n li tch t, mt s t bin bao gm lch khung v chui kt thc, ngn chn 2 m ho
protein bt k phng din no. Do , 2 ca th l mt v d ca gen gi. Gen gi c biu th theo
ch Hylp l , v vy gen ny c k hiu l 2. Loi gen gi ny c xem l kt qu tin ho ca
mt gen c khi biu hin nhng by gi li l gen t bin ln. C mt vi v d khc ca cc gen gi
loi ny, nh cc gen tng t - tubulin ngi, cc gen rARN 5S Xenopus.
Hnh 7.5. S sp xp cc gen tng t
Gen gi 2 ca th cng cha intron v exon lin quan mt thit vi gen cu trc 1. Nhng vic
mt mt cp base codon th 20 ca n gy nn hin tng t bin lch khung lm chm dt ngay
s dch m sau . Gen gi cng c th c to ra khi 2 intron c xen gia l mt exon v khng c
intron no c ct ra khi exon khi gen phin m.
7.3. CC MC IU HO BIU HIN GEN
t bo nhn nguyn thu, cng nh t bo nhn tht, cc c ch iu ho s biu hin ca gen

c th tc ng mt hay nhiu mc khc nhau. S iu ho c th mc ngay bn thn gen,
bng s kim sot thi gian v tc phin m. Cc c ch khc c th hot ng lc dch m hoc sau
dch m (hnh 7.6).
7.3.1. Mc cht nhim sc
Ngay trn si nhim sc c th thc hin cc kiu:
DNaseI ct ADN b gen mt s vng lm tho xon cho cc gen biu hin (1) ca hnh 7.6.
Hai vng c lu : nhy cm v tng nhy cm. Cc vng nhy cm c lin quan n cc gen c
hot tnh cao v nhng gen trc y biu hin (gen hot ng phi).
- ADN Z l dng cu trc siu xon c th lin quan n ng m gen (2).
- S Metyl ho cc base (3) t bo nhn nguyn thu xy ra A v C, cn t bo nhn tht C

v tr 5': S metyl ho lm gen ngng hot ng. V d, nhim sc th X bt hot ng ngi thuc
loi siu metyl ho.
- S thay i cu hnh c th nh hng n s biu hin gen.
Hnh 7.6. S v cc mc iu ho khc nhau
7.3.2. Mc phin m
y l s iu ho nh hng trc tip n vic m hoc ng gen. Kiu iu ho ny thng gp

trong iu ho trao i cht, cng nh trong bit ho t bo.
- S tc ng ca trnh t cis (cng pha) nm trn cng mch ADN nh enhancer (on tng
cng) (4).
- iu ho bi cc yu t trans (tri pha) do cc yu t khng nm cng trn mt mch ADN (5).
- Chn la promoter thch hp (6).
- iu ho suy gim (Attenuation).
7.3.3. Mc sau phin m
iu ho khi mARN ct b cc intron v gn cc exon li vi nhau to mARN trng thnh. Nh

vy, cc h thng nh hng n s trng thnh ca mARN c th kim tra gin tip biu hin ca gen
tng ng. Cc mARN ca t bo nhn tht cn c nhng on khng m ho lin quan ti thi gian tn
ti v ra khi nhn i vo t bo cht.
- Splicing (ct ni) khc nhau (7).
- im polyadenin ho khc nhau.
- t bin trn phn t mARN.
- Bn chu k phn hu ca mARN (8).
- S bo tn cc ARN trong t bo (9).
7.3.4. Mc dch m
S bin i ca cc yu t khi u IF (10).
7.3.5. Mc sau dch m
Sau khi mch polypeptid c tng hp, cc protein nhiu khi chu cc bin i th cp trc khi
biu hin hot tnh. V d, trypsin l enzym phn gii protein trong d dy ch c c hot tnh sau khi
tin cht ca n (proenzym) khng c hot tnh, b ct mt mt on polypeptide.
Cc protein c th chu nhng bin i lp th (allosteric) nh s kt hp cc enzym vi mt s sn

phm c bit c th lm thay i cu trc khng gian ca chng lm cho n c hot tnh.
- Glycosyl ho, phosphoryl ho... (11): gn thm cc nhm cht nh ng, phosphat protein c
hot tnh.
- Tn hiu peptide (12) l on gm khong 20 acid amin nm gn pha u N ca polypeptid, c vai

tr gn polypeptid v ribosome ang c tng hp mch ny vi li ni cht. Trong b my Golgi,
polypeptid c phng thch ra ngoi.
- S phng thch ra protein c hot tnh t mt phc hp nh t proinsulin thnh insulin.
7.4. IU HO HOT TNH GEN CA T BO NHN THT
Trong cc t bo nhn tht c mt s cc im, m s biu hin gen c th c iu ho (hnh

7.7).
Hnh 7.7. Cc im ca gen t bo nhn tht c th iu ho biu hin
Mt s c im ca iu ho hot ng gen t bo nhn tht cn c nhn mnh:
- cc operon ca t bo nhn nguyn thu, cc gen iu ho v cc promoter thng nm gn

nhau, nhng t bo nhn tht th cc gen iu ho t khi nm gn cc promoter do chng kim sot.
- Cc enhancer l nhng trnh t cng nm trn mt phn t vi cc promoter c th c hng trm

cp base pha trc hoc pha sau ptomoter m chng kch thch.
- Trnh t iu ho 5' pha trc cc promoter t bo nhn tht thng rt di, c khi hng chc
kb.
- C nhiu kiu iu ho dng cc yu t c tc ng trans l cc protein.
- S phin m c th c kch thch bi cc tn hiu khc nhau. S iu ho hot ng cc gen t
bo nhn nguyn thu phn ln p li tn hiu ngoi sinh. Ngc li, phn ln s iu ho t bo
nhn tht l p li cc tn hiu ni sinh.
7.4.1. Cc promoter
Tng t nh t bo nhn nguyn thu, cc promoter ca t bo nhn tht cng nm pha trc
im xut pht ca mARN v cng c nhng trnh t chung trong tin ho. Hp TATA nh hng cho
ARN polymerase bt u phin m, pha ngc chiu vi chiu phin m khong 30 bp ADN ng
vt c v v 60 n 120 bp ADN nm men. Hp TATA hot ng c hiu qu cng vi hai trnh t
tng ng pha trc khong 40 bp l CCAAT v 110 bp l trnh t giu GC (hnh 7.8).
Hnh 7.8. Trnh t nucleotide ca promoter

(a) T bo nhn tht; (b) T bo nhn nguyn thu
S thay i hp TATA lm gim tc phin m. S gim tc phin m c o bng s thay

i ca tng base trong promoter. Cc thay i base ngoi hp TATA v cc trnh t pha trc khng
gy tc ng i vi s phin m. Khc vi promoter ca t bo nhn nguyn thu, cc promoter ca t
bo nhn tht kh m bo s nhn bit cc tn hiu mt cch y ARN polymesare khi u
phin m in vivo. Do , xo b hp TATA khng hon ton lm hu b s biu hin gen m lm thay
i v tr bt u phin m.
Hp TATA v cc trnh t pha trc phi c nhn bit bi cc protein iu ho. Chnh cc
protein ny gn vi cc im nht nh trn hp TATA v hot ho s phin m.
Hp TATA c hu ht cc gen t bo nhn tht. Tuy nhin, hu ht cc gen m ho cho cc

protein c bn nng thp (low - abundance housekeeping protein) li khng c hp TATA. Hai trnh
t chung khc c tm thy nhiu gen l: hp CCAAT (pht m l "cat") thng thy t bo
nhn tht bc cao nh - globine th trng thnh, ngi ta tm thy khong 70 bp ngc chiu vi v
tr bt u phin m v c trnh t chung l CCAAT. S khim khuyt hp CCAAT lm gim phn ln
t l phin m, nhng khng lm thay i v tr khi u phin m.
Trnh t th hai c bit n l hp GC hay hp Sp1, cng hin din ngc chiu mt s cc gen
t bo nhn tht, thng l trong cc bn sao nhiu ln. V d, trnh t promoter ca gen Thymidine
kinase c 2 hp GC ngc chiu vi hp TATA (hnh 7.9).
Hnh 7.9. Promoter ca Thymidine kinase c cha cc trnh t tng cng

khc nhau
Hp GC ny cng c kiu hnh tri, c tm thy promoter ca cc gen bo v khng c hp

TATA. Cng nh hp CCAAT, hp GC cn thit cho sao m.
ng lu l, c 2 hp CCAAT v GC hot ng theo c 2 hng. Hp GC tng tc vi mt

protein c gi l Sp1, cn hp CCAAT tng tc vi protein k hiu l CTF (CTF = CCAAT binding
Trascription Factor). N l yu t iu ho trans gn trn hp CCAAT v c th l vi cc protein khc.
7.4.2. Cc yu t tng cng (Enhancer)
Enhancer l cc trnh t c tc ng cis (cng pha), chng tng tc phin m ng k t ngay cc

promoter nm ngay trn cng mt phn t ADN. Tnh c o ca enhancer l ch chng c kh nng
thc hin tc ng cch xa n vi nghn cp base. Ngoi ra, chng c th hot ng bt k hng
no, d pha trc hay pha sau promoter.
Trnh t tng cng u tin c m t virus SV40, c nh v khong 200 bp ngc chiu vi
v tr bt u phin m ca gen sm. Nu loi b trnh t ny s lm gim ng k kh nng sao m t
trnh t promoter ca gen sm. Tuy nhin, s ti chn ca trnh t tng cng vo bt k ni no ca b
gen SV40 s gi li mc biu hin bnh thng. Hn th na, nu trnh t tng cng SV40 c ni
vi gen - globine th s t ngay s biu hin cch xa 200 bp pha trc hay pha sau promoter -
globine v ngay c khi trnh t khuch i SV40 nm cch v tr khi u phin m ti vi nghn cp
base. Gn y, ngi ta tm thy cc trnh t tng cng c cc gen chui nng globine min dch
ng vt c v v gen albumine ca chut. Khong cch gia mt trnh t tng cng v mt trnh t
khi ng ln nht gp gen albumine ca chut l 10 kb.
Xt mt s mt no , cc enhancer tng t cc promoter. Chng c t chc thnh mt dy cc

trnh t c tc ng cis nhn bit cc yu t tc ng (hnh 7.10).
Hnh 7.10. S tng tc ca hormon steroid vi enhancer
Hormon glucocorticoide gn vo protein th th ho tan trong phc hp ny sau li gn vo

enhancer kch thch phin m.
Cc trnh t ti cis c c im chung l chng thng c cu trc gm hai phn i xng nhau. V
d, trnh t p ng vi hormon tuyn gip (TRE - Thyroid hormon Respone Element) di y:
S d nh vy l v cc trnh t ny thng tip nhn cc protein iu ho (yu t trans) di dng

dime (cu to t hai tiu n v).
7.4.3. Cc yu t protein tham gia vo qu trnh iu ho biu hin gen, cc yu t trans
c im chung ca cc yu t trans l gm t nht 2 vng cu trc - chc nng chnh:
- Vng gn yu t trans vo ADN.
- Vng tc ng ln s phin m.
Cc vng cu trc - chc nng ny c lp vi nhau.
Tin hnh ghp hai trnh t ADN c ngun gc khc nhau: vng tc ng phin m ly t mt yu
t trans ng vt c v v vng gn vo ADN ly t yu t trans nm men. Kt qu to ra mt yu
t "lai" c kh nng hot ho cc gen ch tng ng nm men. Gen ch l gen mang trnh t tip
nhn yu t trans. Ngoi hai vng k trn, nhiu yu t trans cn mang mt s vng khc nh: vng
gn hormon, cc ion... (hnh 7.11).
Hnh 7.11. Cu trc mt yu t trans
Hnh 7.11 cho thy cu trc ca mt th th tuyn gip (THR - Thyroid Hormon Receptor) bao gm
nhiu vng cu trc v chc nng, trong hai vng c bit r nht l:
- C: vng gn ln ADN.
- E: vng tip nhn hormon tuyn gip.
Hormon tuyn gip khi gn ln vng E ca th th s lm bin i cu hnh th th. Lc , vng C

ca th th c th gn ln vng ADN iu ho ca cc gen ch v kch thch s phin m ca cc gen
ny.
Cc kt qu nghin cu cho thy cc yu t trans c th phn vo bn nhm cu trc:
- Ngn tay km (zinc - finger).
- Xon - vng - xon (helix - turn - helix).
- Xon - cun - xon (helix - loop - helix).
- Dy ko leucine (leucine - zipper).
Nh vy, cc gen t bo nhn tht c hot ho bi hai trnh t ADN c tc ng cis l promoter
v enhancer. Chng c nhn bit bi cc yu t protein c tc ng trans. Cc yu t ny cho php
ARN polymerase khi s phin m v t tc phin m ti a (hnh 7.12).
Hnh 7.12. C ch "tip xc trc tip" ca trnh t enhancer
7.4.4. Hormon
Hormon l mt trong cc cht iu ho ni ti ca hot tnh gen. l nhng cht c to ra do

mt lot t bo c tc ng n cc t bo khc. Cc hormon ch tc ng n cc t bo c th th
tng ng. Cu trc th th bao gm nhiu vng trong c 2 vng c bit r nht:
- Vng gn vo ADN vi hai cu trc "ngn tay km" (zinc finger).
- Vng gn vo hormon.
S tng tc hormon vi th th gy ra tn hiu tc ng n cc vng c hiu ca ADN, lm hot

ho gen hoc nhm gen tng ng.
- Cc hormon c th kch thch phin m bi mt trong cc c ch sau:
- Tch ADN khi histone v to iu kin cho ARN polymesare bt u phin m.
- Tc ng nh mt cht cm ng, gy bt hot phn t c ch.
- Gn trc tip vi on ADN c hiu, to thun li cho ARN polymesare gn vo phin m.
- Hot ho protein hiu ng, to thnh phc hp hot ho, kch thch ARN polymesare gn vo
phin m.
- Gn vi protein to phc hp hot ho, lm cho ARN polymesare gn vo phin m.
7.5. S KIM SOT CC CHT THNG GP TRONG NHN
Mt s nhm phn t hin din nhiu trong t bo nhn tht nh histone, cc thnh phn ca b my
dch m, cc thnh phn ca mng, t bo,... duy tr s lng ln chng trong t bo phi c cc
chng trnh:
- Phin m lin tc v lp li trong chu trnh t bo.
- Lp li cc gen.
- Khuch i ngoi nhim sc th ca cc trnh t gen c hiu.
7.5.1. S di do ARN
Khng phi tt c cc gen u biu hin cng mt mc . iu ny dn n s khc nhau ng

k v mc tp trung ca cc mARN v cc protein ring l. mARN c phn chia mt cch gng
p thnh 3 nhm: d t, trung bnh v nhiu. Mt t bo n nh c di 100 mARN d, mt vi t bo
ch c 1 hoc 2 mARN d. Trong mt t bo, trung bnh c khong 33% tng s mARN l mARN d
nhiu, mi mARN d ny c th chim ti 1% tr ln trong tng s mARN ca t bo.
Kiu hnh ca mt t bo l thuc rt nhiu vo qu trnh tng hp ca mt hoc vi protein d, v

vy cn phi c cc phn t mARN d. V d, globin trong t bo li ni cht, si c v myosin trong
cc t bo c. V vy, cc mARN d nhiu thng l cc bn sao ca cc gen chuyn bit ca t bo.
Bng 7.3. Cc nhm mARN d vi trng g v gan chut nht
% ca tng mARN trong S lng cc loi

Loi t bo Nhm d
nhm mARN khc
Nhiu 50 1
Vi trng g Trung bnh 15 7
t 35 12.500
Nhiu 22 9
Gan chut nht Trung bnh 41 700
t 37 11.500
Mt t bo cha vi trm mARN d trung bnh, chim khong 33% tng s mARN t bo, nhng
mi loi mARN d ch chim 0,1 - 0,5% tng s mARN. Phn mARN cn li ca t bo gm cc bn
m sao d t. C khong 10000 bn m sao nh vy trong mt t bo in hnh, mi bn sao i din
cho t hn 0,01% tng s mARN.
Cc nhm gen d t hay d trung bnh u nhy cm khng km so vi d nhiu.
Cc loi ARN khc cng c hin tng d. Xenopus c vng t chc hch nhn cha 450 bn sao
ca ADN m ho cho rARN 18S v 28S. Ngc li, trong mi nhn c t 20000 bn m sao ca cc gen
m ho cho rARN 5S v cc gen ny khng nm trong vng t chc hch nhn.
Cc gen m ho cho histone cng c nhiu bn sao c lp li hng trm ln.
S di do ca cc gen nu trn m bo s lng cn thit cho dch m khi cn phi tng hp

hng triu phn t protein.
7.5.2. S khuch i gen
Mt v d v khuch i gen trong nhn l cc ch phnh (puff) ca nhim sc th khng l hay a

si tuyn nc bt Drosophilla. cc ch phnh ny, ADN nguyn nhim sc c khuch i
khong nghn ln.
Trong nhn ca t bo trng Xenopus c ti hng trm nhn con ngoi nhim sc th vi kch thc
khc nhau. Mi nhn con cha cc vng rARN c kch thc khc nhau, vai tr cha r. Cc ADN vng
ny sn sinh nhiu rARN lp rp thnh nhiu ribosome.
Tm li, s biu hin ca cc gen t bo nhn tht rt phc tp. Tuy vy, hng nghn promoter
c iu ho to ra cc mARN mc hp l.
Tm tt
Kch thc v cu trc b gen t bo nhn tht phc tp hn nhiu so vi b gen ca t bo nhn
nguyn thu. ADN ca b gen t bo nhn tht c mc lp li cao, trung bnh v n c.
S iu ho biu hin gen c thc hin t bo nhn tht nhiu mc khc nhau:
- mc nhim sc th, phin m, sau phin m v dch m. B my di truyn khng nhng c

cc gen cu trc m cn c nhng trnh t khc tham gia biu hin gen.
- t bo nhn tht c nhiu c ch iu ho hot ng gen phc tp nh: cc yu t cis (enhancer),

cc yu t trans, cc hormon. Ngoi ra, cn c s d di do ca mt s bn m sao v s khuch i
mt s gen cn thit cho hot ng ca t bo.
CU HI
1. Gi tr C l:
a) Tng s nucleotid c trong t bo
b) Kch thc nhim sc th
c) Kch thc gen
d) Kch thc b gen lng bi
e) Kch thc b gen n bi
2. Nghch l gi tr C
a) L gi tr nghch o ca gi tr C
b) S khc bit gia gi tr C ca sinh vt bc cao vi vi khun
c) S khc bit gia gi tr C ca ngi vi sinh vt bc cao khc
d) S khng tng xng gia kch thc b gen vi s gen
e) S khng tng xng gia b gen n bi v lng bi
3. Cc trnh t lp li ADN t bo nhn tht gm
a) SINE b) LINE
c) CEN d) TEL
e) Tt c
4. Trnh t TEL
a) Thuc nhm telomer b) Bo v u mt nhim sc th
c) Kim hm s tin ho d) Khng gn vi mng nhn
e) a v b
5. Trnh t SINE
a) Cn gi l Alu b) Khng cha nhm Alu
c) Ngn hn LINE d) Di hn LINE
e) a v d
6. Gen gi l:
a) Trnh t lp li thp b) Trnh t lp li cao
c) Trnh t c nht d) Khng m ho cho protein
e) c v d
7. Cc hormon c th kch thch phin m bng cch:
a) Tch ADN khi histone
b) Tc ng nh mt cht cm ng
c) Hot ho protein hiu ng
d) Gn vi protein to phc hp hot ho
e) Tt c
8. Cc mc iu ho biu hin gen t bo nhn tht
a) Cht nhim sc b) Phin m
c) Hu phin m d) Dch m v hu dch m
e) Tt c
9. Enhancer
a) C tc ng cis
b) Hot ng theo kiu trans
c) C kh nng thc hin tc ng cch xa n vi nghn cp base
d) Hot ng theo hng 5 3
e) a v c
10. iu ho hot ng gen t bo nhn tht khc t bo nhn nguyn thu
a) Trnh t iu ho 5' thng rt di
b) Trnh t iu ho 3' thng rt di
c) Ch yu p ng li tn hiu ngoi sinh
d) Ch yu p ng li tn hiu ni sinh
e) a v d
Bi 8
T BIN GEN
MC TIU
- Trnh by c t bin xy ra nh th no, c th c hu qu g vi t bo.
- Nu c cch t bo sa cha ADN h hng trnh t bin.
- Nhn din c mt s bnh di truyn ngi do t bin gy ra.
- Trnh by c tc ng ca cc yu t gy t bin.
- M t c vic s dng cc yu t gy t bin trong phng th nghim v cc phng php

chn lc t bin vi sinh vt.
8.1. M U
t bin theo ngha rng ch cc bin i di truyn xy ra t ngt. Mt t bin gen l mt s thay
i trong trnh t nucleotid trn gen, dn n kt qu l to ra mt protein t bin vi trnh t acid amin
thay i hay tc ng n qu trnh iu khin s tng hp ca cc sn phm gen. Nu protein t bin
c chc nng khc vi dng t nhin th s to ra mt thay i tng ng nhng c tnh c th quan
st c v to ra th t bin. S khc bit c th do hot tnh hay tnh n nh ca enzym. Mt s t
bin c th tn ti trong qun th bn cnh cc dng t nhin lm thnh mt th a hnh. Trong th a
hnh, khng th phn bit gia dng t bin v dng t nhin.
t bin xy ra cc t bo khng sinh sn c gi l t bin sinh dng v to ra cc thay i

c tnh cc b, khng di truyn c nh cc u sc t ngoi da ngi, to ra nt rui. Nhng t bin
sinh dng cng c th l nguyn nhn gy ra ung th v c th l nguyn nhn ca s lo ho.
t bin xy ra trong giao t c gi l t bin dng mm v c truyn cho i sau. Khi

nghin cu cc t bin giao t v o t l t bin sinh vt a bo ni chung v ngi ni ring cn
xt n tnh lng bi. Hu ht cc t bin gen l t bin ln v s khng c pht hin nu hp t
khng c hai bn ca allel t bin. Do vic pht hin v o tn s t bin phi da vo quan st cc
t bin tri ang xy ra trn nhim sc th thng, cc t bin ln v tri trn nhim sc th X. V d,
chng lon sn sn xy ra ri rc l bi cc t bin mi trong gen ca th th yu t tng trng
nguyn bo si. Trong mt nm c by tr s sinh b lon sn sn trn tng s 242 257 tr s sinh, t l
l 7/242 257 1/2 (2 allel/hp t) = 1,410-5.
ngi, cc gen c t l t bin cao (khong 10-4) l bnh u x thn kinh (NF) v bnh lon dng
c Duchenne (DMD), c t l t bin thp (khong 10-6) l gen bnh Huntington. S cch bit hng
trm ln cho thy t l t bin gen c th khc nhau tht s.
C hai gii thch cho s khc nhau v t l t bin ca cc gen:
- Kch thc ch: gen NF v DMD c vng m ho protein rt ln, c nhiu base c th b thay i
hoc lm mt chc nng ca gen.
- im nng: mt s gen nm trong vng nhim sc th nhy cm hn vi s ph hu/ thay i di

truyn hoc gen c cha cc trnh t d b thay i bi cc t bin t pht hn. V d, gen chng lon
sn sn c cha im nng (trnh t CpG).
Trong t nhin, d trong iu kin no, tt c cc gen u c t bin c gi l t bin t nhin

hay t bin t pht. Cc t bin t nhin thng xut hin rt t. Cc gen khc nhau ca cng mt sinh
vt c th c tn s t bin khc nhau nhng tn s t bin t nhin i vi mi gen l n nh.
Tn s t bin c nh gi cn c trn mt ln sao chp, mt ln phn bo hay trn mt giao t

v trn mt t bo trong mt th h (bng 8.1).
Bng 8.1. Tn s t bin t nhin ca mt s gen
Sinh vt t bin Tn s Cn c nh gi
Thc khun th Km hm to r r + 10-8 t bin gen/ ln sao chp

T2
E. coli Ln men Lactose Lac Lac + 2.10-7 T bo t bin/ln phn bo
Leu Leu+ 7.10-7 t bin / t bo
Arg+ Arg 4.10-9 t bin / t bo
Trp+ Trp 6.10-8 t bin / t bo
Ara+ Ara 2.10-6 t bin / t bo
StrS StrR (nhy khng Str) 4.10-10 t bin / t bo
Neuospora Adenin: ade ade + 4.10-8 t bin / bo t v tnh

crassa
Bp R r ( trng) 4,92.10-8 t bin / giao t
C c (mu khng mu) t bin / giao t
Wx wx (t np) t bin / giao t
Y y (vng trng) 2,2.10-6 t bin / giao t
Drosophila Ww 10-5 t bin / giao t
e+ e (mu en h phch) 2.10-5 t bin / giao t
ey+ ey (c mt khng mt) 6.10-5 t bin / giao t
Tn s t bin c th hiu mt cch o ngc. V d, rui dm thn xm bnh thng (e+) t bin
thnh thn c mu en h phch e (ebony) vi tn s 2.10-5, ngha l trong 105 giao t c hai t bin t
e+ e. Hoc v d E. coli t bin nhy cm vi streptomycin thnh khng streptomycin vi tn s
4.1010 t bin trong mt t bo mt th h. d hiu ta c th tnh ngc li, tc l trong 10 t t
bo ca mt th h c 4 t bin StrR xut hin ngu nhin.
Tuy tn s t bin ca tng gen l rt thp, nhng tng cc t bin ca nhiu gen l mt s ng
k, c ngha quan trng trong tin ho. Cc kt qu nghin cu in vivo v in vitro trn t bo ngi cho
thy t l t bin ngi khong 106/gen/th h. Cc vi sinh vt t bo nhn nguyn thu v t bo
nhn tht cng c t l t bin tng t. C th c tnh cc thay i xy ra trong mi th h da vo t
l t bin ngi nh sau: 106 t bin/gen x 5.104 gen/b gen n bi bng 5.102 t bin trn giao t
(5/100 hoc 1/20). Mi hp t c (1/20).2 giao t trn hp t bng 1/10 c hi mang t bin mi u
trong b gen. Con s ny c v rt cao nhng cn nh rng hu ht cc t bin l ln v nh vy s
khng c biu hin trong iu kin d hp t.
S thay i cc base c th xy ra trong vng khng m ho ca phn t ADN v cc acid amin b

thay i cng c th khng phi lc no cng lm thay i c tnh protein. Do vy, thay i trnh t
nucleotid c th khng biu hin thnh s khc bit tnh trng v do khng phi l i tng ca
chn lc. Cc t bin ln tch t trong b gen.
Hu ht cc t bin l bt li v do cc c ch c hiu thng c dng gim thiu cc

bin i trong chui ADN hoc phc hi trnh t gc bng cch sa cha cc t bin.
Trong phng th nghim, t bin cung cp cng c c gi tr cho vic nghin cu s sp xp cc

gen, chc nng ca gen v cch kim sot hot ng ca chng. Thc hin iu ny vi sinh vt d hn
l sinh vt bc cao.
K thut cm ng t bin c s dng trong cy trng v vt nui to ra cc chng mi v

trong cng ngh vi sinh hc hin i to ra cc chng vi sinh vt lm tng nng sut v to ra sn
phm mi.
K thut ti t hp ADN cho php chn mt t bin c hiu vo mt gen quy nh trc (t bin
im nh hng) c th to ra cc t bin tt v hu ch.
8.2. CC LOI T BIN
8.2.1. t bin im
t bin im l kt qu ca thay i mt base trong chui ADN gy bi tc nhn gy t bin hoc

sai st trong sao chp ADN.
8.2.1.1. Thay th cp nucleotid
Ph bin nht l chuyn v, pyrimidin c thay th bi pyrimidin khc hay purin bi purin khc (A
G, C T). S chuyn v, bt cp sai, c th gy bi acid nitr hoc cc ng ng base nh 5 -
bromo - 2 - deoxyuridin (BrdU). Cch t bin im t ph bin hn l o chuyn, pyrimidin c thay
th bi purin hay purin bi pyrimidin (C/T A/G). t bin im c khuynh hng c trng l t
bin li (hi bin), ngha l chuyn li dng nguyn thu. iu ny xy ra v ln t bin tip theo ti
cng mt im c mt phn ba c hi quay li chui ban u. C ba loi t bin im tu theo codon
mang t bin m ho:
t bin ln hay cn gi l t bin cng ngha: m ho cho cng acid amin, khng c nh hng
n protein cui. V d, codon GCA hoc GCG trong mARN u ngha l arginin (loi ny thng ng
vi t bin chuyn v v tr th ba - base "linh hot" ca codon).
t bin lch ngha: m ho cho acid amin khc, c th lm protein khng c chc nng. V d,
trong chui protein - globin gy bnh thiu mu hng cu hnh lim, GAG m ho cho glutamat; nu
thay bng GUG s m ho cho valin. t bin lch ngha c th gy hu qu rt nghim trng nh thiu
mu t bo li lim, hoc nh nh hemoglobin C (acid amin v tr 6 ca - globin b thay th bi acid
amin khc) hoc khng c kiu hnh.
t bin v ngha: m ho cho codon kt thc, c th lm ct protein. Tc ng ca cc t bin
v ngha tu thuc protein b ct bao nhiu v mc cn cho hot ng ca protein.
t bin thay th base c th xy ra trong promoter hoc cc vng iu ho 5 ca gen hoc trong
intron v c th nh hng n s phin m, dch m hoc mc xon bn ca ADN. Nhiu -
thalassemia l hu qu ca cc t bin khng cu trc ny nh hng mc biu hin cc gen globin.
Mc nh hng tu thuc nucleotid b thay th v v tr ca n trong ADN.
Hnh 8.1. Cc loi t bin im
8.2.1.2. t bin lch khung
Cc t bin ny do chn hoc mt mt hoc nhiu nucleotid trong vng m ho ca gen. iu ny

lm thay i khung c, v cc codon l cc nhm ba nucleotid nn c th c ba khung c nhng ch
mt ci c dng. t bin loi ny thay i tt c acid amin pha sau t bin v rt d to sn phm
khng chc nng v khc hn vi protein bnh thng. Ngoi ra, cc khung c sai thng cha cc
codon stop, s lm qu trnh tng hp protein dng sm.
Vic thm hoc bt mt nucleotid vo khung c s lm thay i acid amin k t tr i. Nu t

bin thm hoc bt cng xy ra trn mt phn t ADN th do trao i cho gia cc base tng ng m
t bin m c th sa cha li khung c sai do t bin dng gy ra. Nh ch c acid amin no
nm gia hai im t bin b sai m thi. Nh vy, t bin m tc ng nh mt t bin km hm
ngoi gen i vi t bin dng hoc ngc li.
8.2.2. t bin a im
Cc t bin a im l nhng thay i tc ng n hn mt base, thay i t hai n hng ngn

base, nhng thng nht l ch vi base. Hu qu thng l xo b mt phn ca trnh t m ho. t
bin a im khng c tnh hi bin, ngha l t bin c n nh.
S xo mt on ln s lm thay i hon ton protein to ra, thm ch xo b on ngn cng gy

hu qu nghim trng nu n lm thay i khung c, ngha l nu s lng base b xo khng chia ht
cho ba.
S chn a im c th xy ra do gen nhy hoc cc li khi sao chp ca cc yu t lp li (v d cc

lp li AT). Hu ht khi gen b chn s b lch khung hoc thay i kt ni trong mARN, c hai trng
hp ny u lm thay i sn phm ca gen. Gen nhy di ti vi ngn base (kb) v v vy, n lm
ngng qu trnh sao m v dch m ca bt k gen no chng xen vo. Gen nhy c hai loi,
retrotransposon v transposon. Cc transposon c th di chuyn trc tip t v tr ny sang v tr khc
trong b gen, retrotransposon trc ht c phin m thnh ARN v sau tr li thnh ADN bi
enzym phin m ngc v chn li vo b gen. Transposon rt c ch cho cc nh nghin cu lm

cng c bin i ADN trong c th sng.
8.3. NGUYN NHN T BIN
C hai loi t bin l t bin t nhin (xy ra mt cch t pht) v t bin cm ng, gy ra bi
cc tc nhn t bin.
8.3.1. t bin t nhin
Cc t bin xy ra trong t nhin mt cch ngu nhin vi tn s nht nh v khng xc nh c

ngun gc. Bt k mt tin trnh no dn n s gn sai base trong qu trnh sao chp ADN s to ra
mt t bin im. Tn sut sai st t nhin vo khong 10-10 base. T l ny thp t c nh kh
nng sa li ca cc ADN polymerase ch yu. Cc enzym ny ko di ADN theo hng 5' 3' nhng
c hot tnh exonuclease 3' 5'. Nu li nm gia base c chn cui cng vi si khun c pht
hin, enzym loi b base sai v thay th base ng vo. Chui m ca mt protein trung bnh 300 acid
amin l khong 1000 base chiu di. T l t bin t pht l mt trong 107 bn m sao ca gen s c
mt t bin im, ngha l 107 t bo s c t bin v gen .
t bin t nhin mc phn t gm c h bin, kh amin, chuyn v, o chuyn, t bin lch

khung, oxy ho ph hu.
- H bin: 4 loi base trong ADN c th tn ti dng h bin, tc l c kh nng tn ti hon

chuyn gia hai dng (Keto Enol v Amino Imino). Base pyrimidin bnh thng c dng keto
(C=O), nhng i khi chng cng c dng enol (C - OH). dng enol, thymin c th bt cp vi guanin
v enol cytosin c kh nng bt cp vi adenin (hnh 8.2a) tng t base purin bnh thng tn ti di
dng amino (NH2) nhng cng c dng h bin imino (=NH) him gp, c th dn n s bt cp nhm.
Nu trong khi ADN sao chp, G dng enol, polymerase s thm T vo thay v bnh thng l C v quy
lut bt cp b thay i (khng phi do li ca polymerase). Kt qu l s chuyn v G - C thnh A - T;
s h bin ch gy cc t bin chuyn v.
- Kh amin: Cc phn ng kh amin thng thng l phn ng thu phn cytosin thnh uracil, qu
trnh ny phng thch ammonia v kh amin 5 - methylcytosin thnh thymin v ammonia (hnh 8.2b).
Trong ADN, s kh amin cytosin t pht c chnh li bng cch loi b uracil (v uracil khng c
trong ADN) v thay th bng cytosin, cn s kh amin 5 - methylcytosin khng c chnh v c ch
sa cha khng nhn din Thymin l li. Trong ADN b gen cytosin ti cc trnh t CG (CpG) thng
b methyl ho, iu ny ngha l u CpG xy ra trong gen, n l "im nng" cho t bin. Mi y
ngi ta tm thy trong gen lin quan n lon sn sn c nhng im nng nh vy.
- t bin lch khung (chn hoc mt trn mt si), thng l li ca polymerase khi sao chp cc
on lp li ca mt nucleotid. Mi polymerase c chnh xc khc nhau. Yu t chnh nh hng n
chnh xc ca polymerase l c exonuclease c sa 3 - 5, lm loi b cc base bt cp khng ng
chn vo bi polymerase.
- Oxy ho ph hu do cc gc oxy. Cc gc oxy tng trong t bo do chuyn ho oxy ho (v cng

c to bi cc tc nhn vt l nh tia phng x). Sn phm oxy ho quan trng l 8 - hydroxy guanin
bt cp sai vi A, to o chuyn G - C thnh T - A.
Hnh 8.2. Hiu qu t bin ca s thay i tnh cht cp base b sung

(a) ng phn ca thymin v 5 - bromouracil; (b) s kh amin ho cytosin thnh uracil.
8.3.2. t bin cm ng
Cc tc nhn lm tng tn s t bin cao hn mc t nhin c gi l cc tc nhn gy t bin.

Cc tc nhn ny c th lm thay i cu trc hoc trnh t ADN. Cc tc nhn vt l gy t bin nh
phng x, tia X, tia t ngoi... Nhiu ho cht l cc tc nhn gy t bin nh cc ng ng ca cc
base nit, acid nitr (HNO2), cc cht alkyl ho mnh.
8.3.2.1. Tc nhn ho hc gy t bin
Ngi ta bt u bit n tc ng gy t bin ca cht ho hc khi s dng kh ngt nit trong

Chin tranh th gii ln th nht v hai, cht ny c th gy t bin trong t bo. Sau , nhiu cht
ho hc gy t bin khc c xc nh v c kim sot nghim ngt. C th phn bit cc tc
nhn t bin ho hc da vo kiu tc dng ca chng, mt s cht gy t bin theo c ch tng t
t bin t nhin, mt s khc c c ch ging bc x.
Tc nhn gy t bin ho hc c th c chia ra lm 4 nhm:
- Nhm base ng ng.
- Cc cht ho hc thay i cu trc v c tnh bt cp ca cc base, gm: cc cht kh amin v cc

cht alkyl ho.
- Nhm chn vo ADN (tc nhn lm lch khung).
- Cc tc nhn thay i cu trc ADN.
Base ng ng:
Cc cht ho hc ny c cu trc ho hc ging cc purin v pyrimidin v c th c gn vo ADN

ti v tr ca cc base bnh thng khi sao chp ADN.
Hnh 8.3. Tc ng gy t bin ca bromouracil (BU)
- Bromouracil (BU) l hp cht nhn to dng rng ri trong nghin cu. Ging vi thymin (c
nguyn t Br thay cho nhm methyl) v s gn vo ADN v bt cp vi Adenin ging nh Thymin. C
nhiu kh nng h bin thnh dng enol (BU*) v c kh nng bt cp vi Guanin. Nh vy, BU c th
xm nhp v bt cp b sung vi Adenin thay cho Thymin to A - BU v vng sao chp tip theo BU
li h bin thnh BU* v bt cp vi Guanin to BU* - G, do A - T c thay th thnh G - C (hnh
8.3a). Trong khi BU* c th xm nhp sai, khi gn thay ch cho Cytosine to G - BU*, sau BU*
li h bin thnh BU v tip theo bt cp vi Adenin to A - BU v vng sao chp tip theo Adenin
bt cp vi Thymin thnh A - T (hnh 8.3b).
- Aminopurin l ng ng ca Adenin, c th bt cp vi Thymin hoc (rt t) vi Cytosin, gy

chuyn v A - T thnh G - C hoc G - C thnh A - T. Cc ng ng base gy chuyn v nh gy h bin
t pht.
Cc cht ho hc thay i cu trc v c tnh bt cp ca cc base:
- Cc cht kh amin. V d in hnh: acid nitr to bi s phn gii cc nitrit (cht bo qun)
trong thc phm lm Cytosin thnh Uracil, Methylcytosin thnh Thymin, v Adenin thnh Hypoxanthin
kh amin, Hypoxanthin trong ADN bt cp vi C gy chuyn v. Methylcytosin c mt trong cc gen
ca sinh vt bc cao c th nh hng ti hot tnh phin m. Cc base ngoi nh Xanthin,

Hypoxanthin v Uracil c th khi ng c ch sa cha. Thymin, dn xut t Methylcytosin ni sinh
v nhiu s chuyn v c th xy ra theo cch ny. Cc gc 5 - methylcytosin, sau tr thnh "im
nng" ca s t bin. Hydroxylamin (NH2OH) ch kh amin ca Cytosin, do c tc ng rt gii
hn:
Cc cht alkyl ho. V d in hnh: Nitrosoguanidin (NTG), Methyl Methane Sulfonat (MMS),
Ethyl Methane Sulfonat (EMS) l cc tc nhn ho hc gy t bin phn ng vi cc base v thm cc
nhm Methyl hoc Ethyl nn cn gi l cc tc nhn alkyl ho (bng 8.2). Tu yu t b tc ng, chng
c th gy t bin t nht bng 3 cch:
Thm nhm Methyl (- CH3) hay ethyl (- C2H5) vo Guanin to base ng ng ca Adenin dn
n s bt cp b sung sai (hnh 8.4).
Mt purin do Guanin b alkyl ho to l hng trn ADN, khi sao chp c th lm t mch
(hnh 8.4).
Lin kt cho gia cc mch ca mt hoc cc phn t ADN khc nhau lm mt nucleotid.
Guanin l base c th d b tn cng nht, vng pyrimidin, c bit l cc nhn pyrin d dng b
alkyl ho. Cc v tr ny l cc v tr lin quan trc tip n vic ghp cp nh 6 - keto oxygen ca
guanin hay 4 - keto oxygen ca Thymin v cc v tr cch xa v tr ghp cp nh N - 7 ca Guanin.
Hnh 8.4. Hiu qu d on khi x l Guanin
vi tc nhn alkyl ho (EMS hay MMS)
Bng 8.2. Mt s tc nhn alkyl ho c s dng gy t bin cm ng
Tn ho hc Vit tt Cng thc ho hc
Methyl methan sulphonat MMS CH3 - SO2 - O - CH3
Ethyl methan sulphonat EMS CH3 - SO2 - O - CH2 - CH3
Dimethyl sulphonat DMS CH3 - O - SO2 - O - CH3
Diethyl sulphat DES CH3 - CH2 - O - SO2 - O - CH2 -

CH3
N - methyl - N - nitro - N - nitrosoguanidin MNNG CH3 - N.NO - C:NH - NH - NO2
Nitrogen mustard (kh ngt nit) CH3 - N - (CH2 - CH2 - Cl)2
Sulphur mustard (kh ngt sulphur) S - (CH2 - CH2 - Cl)2
Ethylen oxid EO O - (CH2)2
Ethylen imin EI
Nitrosoguanidin NG
N - ethyl - N - nitrosourea (ENU, cng thc ho hc C3H6N3O2) l tc nhn alkyl ho gy t bin

mnh trn chut. T l t bin ca n l 1 t bin mi ti mt gen ring bit trong 700 giao t. ENU
tc ng bng cch chuyn nhm ethyl ca n vo base nit (ch yu l Thymin) trong acid nucleic.
ch chnh ca n l cc t bo mm nguyn bo tinh, t m tinh dch trng thnh.
Cc tc nhn chn vo ADN (lm lch khung):
Nhm ny gm cc cht Proflavin, cam Acridin, Ethidium bromide dng trong phng th nghim
lm cht nhum. Tt c u l cc phn t a vng phng tng tc vi cc base ca ADN v chn vo
gia chng. S chn ny lm "dn" si i ADN v ADN polymerase b "nh la" chn base vo i
din nhiu hn bnh thng, lm lch khung ADN to thnh.
Cc tc nhn thay i cu trc ADN:
Cc tc nhn ny c th l:
- Cc phn t ln (knh cng) gn vo base trong ADN v lm chng tr thnh khng m ho. V
d, N - acetoxy - 2 - acetylaminofluorene (NAAAF)
- Cc tc nhn gy lin kt cho trong v gia cc si. V d cc psoralen c trong mt s rau v

dng trong iu tr mt s bnh v da.
- Cc cht ho hc gy t si ADN. V d cc peroxid.
- Cc hydrocarbon a vng. V d cc benzypyrene c trong xng.
8.3.2.2. Tc nhn bc x gy t bin
Bc x l tc nhn t bin c bit u tin (nm 1920). Bc x c pht minh vo nhng nm

1890: Roentgen pht hin tia X vo 1895, Becquerel pht hin hot tnh phng x vo 1896 v Marie
vi Pierre Curie pht hin cc nguyn t phng x vo 1898. Ba pht hin ny v nhng ci khc
khai sinh Vt l nguyn t v kin thc v bc x in t.
Tc dng gy t bin ca bc x c 2 c im:
- Khng c ngng tc dng tc l khng c liu lng v hi.
- S lng t bin t l thun vi liu lng phng x, khng ph thuc vo cng v thi gian
chiu x.
Cc bc x in t:
Bc sng ca cc bc x ny thay i tu theo loi bc x v t l nghch vi nng lng ca

chng.
Cc sng di nht (radio AM) c nng lng t nht, trong khi cc sng ngn hn l FM, TV, viba,
hng ngoi, kh kin, tia t ngoi (UV), tia X v tia gamma ln lt c nng lng gia tng. Tia t
ngoi (cn gi tia cc tm) c nng lng phng x cao nn c ngha sinh hc.
Bc x ion ho:
Tia X, tia gama, ht phng x v c th lm ion ho cc phn t trn ng i ca chng, do

c tc ng mnh n cc phn t sinh hc.
Bc x UV khng ion ho nhng c th phn ng vi ADN v cc phn t sinh hc khc, v cng l

tc nhn t bin quan trng.
n v dng cho tt c cc loi bc x ion ho hin nay l rem (roentgen equivalent man = 0,01J/kg
ngi b chiu x), do 1 rem ca bt c bc x ion ho no cng c tc dng sinh hc nh nhau. n
v c dng trc y l rad (radiation absorbed dose). Tuy nhin rad ca cc bc x khc nhau s
khc nhau: 1 rad ca cc ht c tc dng ph hu ln hn 1 rad ca cc tia gamma, ngha l cc ht
c RBE (relative biological effectiveness) ln hn 1 rad ca cc tia gamma. Mi tng quan gia cc
n v ny l: rad RBE = rem.
Ngoi ra, cn xt n loi nng lng v liu bc x ton phn, t l liu: cng s rem, nu chiu tp
trung (t l liu cao) gy bng v ph hu da, trong khi chiu nh trong thi gian di (t l liu thp) s
ch lm tng nguy c t bin v ung th.
Ngun bc x:
Cc ngun bc x t nhin sn xut bc x nn. Ngun bc x t nhin gm c cc tia v tr t Mt

Tri v khng gian, t cc nguyn t hot ng phng x trong t, trn mt t v trong khng kh
(radon). Mc bc x nn thay i tu vng a l.
Ngoi ra, con ngi to ra ngun bc x nhn to, gp phn tng mc bc x. Trong c cc th
nghim y t (v d chn on bng tia X), th ht nhn v cc nh my nng lng ht nhn, cc sn
phm khc (tivi, my d khi, kim tra bng tia X sn bay).
Tc dng sinh hc ca bc x:
Bc x ion ho sn sinh cc gc t do ca nc (gc hydroxyl), gy nhiu h hng i vi t bo v

sinh vt. Cc gc t do c cc in t khng bt cp v rt hot ng v mt ho hc, s tng tc vi
ADN, protein, lipid trong mng t bo,... Nh th tia X c th gy h hng ADN v protein, c th lm
hng c quan, ngn phn chia t bo hoc lm cht t bo. Cc loi t bo phn chia nhanh (t bo to
mu, t bo tu xng, nim mc tiu ho) b nh hng bi bc x ion ho nhiu nht v nghim
trng ca tc ng tu thuc liu nhn.
nh hng di truyn ca bc x:
Bc x ion ho gy nhiu nh hng trn ADN thng qua cc gc t do v tc ng trc tip:
- t mt hoc c hai si (c th dn n sp xp li, xo mt, mt nhim sc th, t bo cht nu

khng sa cha).
- Ph hu cc base (t bin).
- Lin kt cho trong ADN hoc ADN vi protein.
Tia cc tm (UV):
Bc x UV t nng lng v do khng ion ho, nhng nng lng ca n c u tin hp ph

bi cc base ca ADN v bi cc acid amin thm ca cc protein, v th n cng c nh hng sinh hc
v nh hng di truyn quan trng.
UV c phn loi theo bc sng: UV - C (180 - 290 nm) c tnh st trng mnh nht v gy cht,
khng c trong nh sng mt tri v n c hp ph bi tng ozon; UV - B (290 - 320 nm) l phn
on gy cht/t bin chnh ca nh sng mt tri; UV - A (320 nm - kh kin) l "cn UV", cng c
tc dng xo mt (v n to cc gc oxy) nhng n sn xut rt t dimer pyrimidin. Cc tn thng gy
cht chnh l cc dimer pyrimidin trong ADN (to bi UV - B v UV - C), cc dimer ny l lin kt
ng ho tr gia cc pyrimidin k nhau trong mt si. Cc dimer ny lm ngn chn s phin m v sao
chp ADN v gy cht nu khng sa cha. Chng c th kch thch t bin cng nh sp xp li
nhim sc th.
Trong t bo, cc cht hu c c mch vng ch yu nh purin v pyrimidin hp thu trc tip UV.
Tia t ngoi c kh nng xuyn thu thp nn ch tc ng ln cc sinh vt n bo v giao t. ADN hp
thu tia t ngoi mnh nht bc sng 257 nm ( 260 nm), y chnh l bc sng lm tng tn s t
bin ht phn cy bp (ng). Di tc ng ca tia t ngoi, cytosin gn thm phn t nc vo lin
kt C=C ca mch vng (Hnh 8.5) v thymin b t lin kt C=C mch vng, ni 2 phn t thnh
thymin dimer.
Stone v cc cng s nhn thy tn s t bin tng ln Staphylococcus aureus khi mi trng
nui chng c chiu tia UV trong thi gian ngn trc khi cy vo. y l tc ng gin tip ca tia
t ngoi.
Hin tng quang phc hi l mt c trng trong tc ng ca tia UV. Sau khi chiu tia t ngoi
ln t bo, nu ngoi nh sng, th cc sai hng phn ln c phc hi. nh sng c tc ng hot
ho enzym sa sai, ct t cc thymin dimer.
Hnh 8.5. Tc ng ca tia t ngoi ln cc pyrimidin

(a) Dimer thymin - cytosin (b) Dimer thymin - thymin
8.4. CC C CH CHNG LI T BIN
V s sai hng ADN xy ra t pht v l hu qu ca cc tc nhn mi trng mi ni nn hu ht

cc sinh vt c mt vi kh nng sa cha ADN ca chng v ADN l i phn t duy nht c sa
cha bi t bo. C th chia cc c ch "sa cha" thnh ba loi:
- o nghch sai hng: c ch ny n gin nht, enzym hot ng phc hi cu trc bnh thng
khng c gy khung.
- Loi b sai hng: ct b v thay th base hoc phn nucleotid sai hng hoc khng thch hp.
- Dung np sai hng: khng sa cha tht s, chp sai hng v tip tc sng.
8.4.1. o nghch sai hng
8.4.1.1. Quang phc hi
y l h thng sa cha n gin nht v c l xa nht. C s tham gia ca mt enzym c kh

nng ct cc dimer pyrimidin (lm t cc lin kt ng ho tr) khi c nh sng (hnh 8.6).
Enzym photolyase xc tc phn ng ny, n c nhiu trong vi khun, t bo nhn tht bc thp, cn
trng v thc vt. Khng c trong t bo ng vt. Gen (phr) c trong ng vt c v nhng m ho cho
protein c chc nng h tr loi sa cha khc.
Hnh 8.6. Quang phc hi
8.4.1.2. Sa sai bng cch lm mt nhm alkyl (dealkylation)
Sa cha trc tip cc tn thng bng O6 - methylguanin - ADN methyltranferase. Enzym ny

chuyn nhm alkyl t nguyn t oxi trn ADN sang gc Cystein trn enzym v phn ng khng thun
nghch v enzym b mt hot tnh (hnh 8.7). Cc li c sa cha gm O6 - methylguanin, O4 -
methylthymin, v cc methylphosphotriester gy ra bi cc tc nhn alkyl ho nh N - methyl - N -
nitro - N - nitrosoguanidine (MNNG). Enzym ny cng sa cha cc guanin b alkyl ho khc nh O6 -
ethylguanin v O6 - butyl - guanin, mc d hiu sut thp hn nhiu. Cc MTase c tm thy tt c
sinh vt c nghin cu.
E. coli gen m ho cho enzym ny (39 kDa) l ada. V tr Cys69 trn enzym nhn gc metyl t
phosphodiester v thay i cu hnh cho php n hot ho promoter ca cc gen ada, alkA
(methyladenin DNA glycosylase), alkB, v aidB, nh tng kh nng khng vi cc tc nhn alkyl
ho. Trong khi v tr Cys321 nhn gc metyl t O6 - MeGuanin v O4 - MeThymin, khi phc base
bnh thng.
Hnh 8.7. C ch sa sai bng methyltransferase
8.4.1.3. Ni cc ch t ca si n
Tia X v mt s ho cht nh peroxid c th gy ra nhng ch t trn khung ADN. ADN ligase

nhanh chng sa cc ch t n gin trn mt si. Cc vi sinh vt t bin mt ligase c mc ti t hp
cao hn v u ADN sinh ti t hp c hot ho cao. ngi ch mi bit m 46BR c t bin trn
c hai gen ADN ligase I, lm tng trng chm, suy gim min dch v nhy cm vi Mt Tri, cht tr
do u lympho. Cc t bo nguyn bo si ngi c 46B d b dit bi cc tc nhn lm hng ADN,
trong c bc x ion. Ngoi ra, hi chng Bloom cng lin quan vi khim khuyt ADN ligase
(mc d protein ca hi chng Bloom l ADN helicase); t bo nui cy ca bnh nhn ny c mc bin
dng nhim sc th v t bin t pht cao.
8.4.2. Loi b h hng
8.4.2.1. Sa sai bng cch ct base (glycosylase)
Loi b ch sai nh mt hoc vi enzym N - glycosylase. N - glycolase nhn bit base bin i v
thu gii lin kt N - glycosilic ni base vi ng pentose. iu ny to ra mt v tr AP (khng c
purin hoc pirimidin), ngha l c mch ng - phosphat nhng b thiu base nit. V tr AP sau
c ct b bi AP endonuclease v ADN polymerase chp y li da vo khun l mch b sung i
din v c ligase ni lin li.
8.4.2.2. Sa cha lch i
Qu trnh ny xy ra sau sao chp ADN nh l "nh vn kim tra" cui cng s chnh xc ca sao
chp. E. coli, c ch ny lm tng chnh xc ca sao chp ln thm 100 1000 ln. Sa cha ny
c tin hnh bi mt nhm cc protein c th qut ADN v tm cc base bt cp khng chnh xc
(hoc cc base khng bt cp) lm sai lch hai chiu trong xon kp. Nucleotid khng chnh xc phn
di s c loi b v ADN polymerase s hot ng ln th hai c trnh t ng.
Cc protein sa cha lch i ngi mi c xc nh v rt ging t bo nhn nguyn thu

nh E. coli v t bo nhn tht n gin nh nm men.
8.4.2.3. Sa cha bng ct nucleotid - NER
H thng ny lm vic trn "khi" ADN sai hng, lm ngng sao chp ADN v phin m. Qu trnh
gm ct si ADN cha sai hng bi endonuclease pha sai hng, tip theo loi b on ngn cha
vng sai hng bi exonuclease, khong trng to thnh c lp vo bi ADN polymerase (hnh 8.8).
Hnh 8.8. Sa cha bng ct nucleotid
Nhiu sinh vt b t bin khim khuyt NER, d b dit v t bin bi UV v cc ho cht hot
ng ging UV. Ngi c bnh di truyn xeroderma pigmentosum nhy cm vi nh sng mt tri, h
c nguy c cao b ung th da nhng vng c th phi di Mt Tri v c cc khim khuyt trong cc
gen tng ng vi h thng NER t bo nhn tht n gin. Cc t bin NER sinh vt bc thp
nhy vi UV v tng mc t bin v ti t hp cm ng bi UV (v chng khng th dng phng
php NER chnh xc loi b cc dimer pyridin v phi dng cc h thng gy t bin hoc gy ti t
hp).
8.4.3. Dung np ADN sai hng
Khng phi tt c cc ADN sai hng c loi b ngay m mt s c th tn ti mt thi gian. Nu

ti chc ba sao chp ADN c ADN sai hng nh dimer pyridin, qu trnh sao chp s ngng li.
Tuy nhin t bo nhn tht, sao chp khi ng nhiu v tr v c th hi phc li si di ca

dimer, cha li "khong trng" ca si n ADN khng sao chp. Khe ny c th nguy him nu khng
dimer na khi t bo phn chia. Khong trng c th c sa bng ti t hp vi tng ng hoc vi
thanh nhim sc em, to hai phn t con nguyn vn, mt ci vn cn cha dimer.
8.4.3.1. Sa cha bng ti t hp
y l c ch sa cha thc y ti t hp lp khuyt si con khng dimer v l cch chp i

ph tn thng lu tr khng m ho trong ADN (hnh 8.9). Loi ti t hp sa cha ny ni chung
chnh xc (mc d n c th gy tnh ng hp ca cc allel ln c hi) v i hi s tng ng ca si
nhim sc em. Sn phm ca cc gen BRCA1 v BRCA2 nhy cm vi ung th v ngi c th tham
gia sa cha bng ti t hp cng vi tng ng ca cc gen RAD51v RAD52 ca nm men.
Hnh 8.9. Cc bin c trong sa cha bng ti t hp
Loi sa cha bng ti t hp th hai l phn ng lin kt u khng tng ng sa cc u

ADN b t nh gy do bc x ion ho v cc cht ho hc c tc ng gy t bin tng t. H thng
sa cha ny cng c s dng bi cc t bo B v T ca h thng min dch ti sp xp di truyn
cn cho hot ng ca chng. Protein Ku70, Ku80 v cc protein kinase ph thuc ADN cn cho lin
kt u khng tng ng. Cc dng t bo ca loi gm nhm c cc t bin trong cc gen ny rt d
b dit bi bc x ion ho v khim khuyt ti sp xp h thng min dch.
8.4.3.2. Sa cha bng t bin
Mt cch khc ADN polymerase b ngn ti dimer l thay i tnh c hiu ca n, nh c th

chn bt c nucleotid i din dimer no v tip tc sao chp (cch "t bin hoc l cht") (hnh 8.10).
Cch ny xy ra vi khun, ngi ta ngh n c th xy ra t bo nhn tht tuy cha bit r c ch.
y l l do ti sao i khi sa cha c th gy t bin.
Hnh 8.10. Sa cha bng t bin
8.4.4. Sa sai nh h thng SOS (cp cu)
H thng sa cha ny c kch hot nu cc c ch trn b qu ti bi cc t bin lan rng.

Trong trng hp ADN b tn thng ngng sao chp, th phn ng SOS hi phc sao chp v chuyn
sai hng thnh sa sai trong khi nhn i. E.coli quan st s ph hu ADN lm m ra khong 20
gen ca h thng SOS, c kim sot m bi cht km hm lexA. Cht ny gn vo hp SOS chng lp
cc promoter ca cc gen SOS.
Khi c nhiu sai hng cp cu, lexA b kch thch, thay i cu hnh, t ct v mt hot tnh km
hm. Lc , cc gen SOS c m ra. Nu sa sai khng kp, t bo phi chp nhn hoc b t bin
hoc b cht.
8.5. CC TNH TRNG T BIN V PROTEIN T BIN
Mt sinh vt b t bin n gin l mt sinh vt biu hin nhng tnh trng bt thng di truyn
c. Hu ht nhng tnh trng ny c th thy c nh nhng t bin sc t gy nn bnh bch tng,
thay i mu sc bn ngoi ng vt v mu sc hoa bt thng.
Trong y khoa, tm thy cc loi t bin khc nhau ngi. ng dng cc nh lut di truyn
Mendel, ngi ta chng minh rng mt vi bnh ngi c di truyn nh l cc bnh do mt gen
ring l quy nh.
Nm 1908, hn 30 nm trc khi c gi thit "mt gen - mt enzym" ca Beadle v Tatum, nh vt

l ngi Anh Sir Archibald Garrod trnh by mt lot quan nim v d tt ngi. Trong , c mt
d tt c ng gi l "ri lon bm sinh v trao i cht". Garrod cho rng, mt s d tt gy nn do c
th mt kh nng thc hin cc qu trnh ho hc mang bn cht di truyn.
Cc ri lon chuyn ho bm sinh cn c gi l cc bnh phn t v bnh c ngun gc l nhng

sai st mc phn t, nhng biu hin trc tip ca bnh l nhng phn t protein.
Hnh 8.11. Chui phn ng sinh ho ca bnh Alkapton niu v Phenylketon niu,
hai s ri lon di truyn ca s thoi qu acid amin thm
Bnh t bin u tin do Garrod nghin cu l bnh Alkapton niu. Nc tiu ca bnh nhn tr
nn en khi ngoi khng kh. Bnh xy ra do thiu enzym homogentisate oxydase trong gan nn
khng phn hu c acid homogentisic (tn c l alcapton) thnh mt sn phm khng mu l acid
acetoacetic. Lc by gi acid homogentisic tch t li trong nc tiu v c mu en khi gp khng kh.
Lin quan ti ri lon chuyn ho bm sinh, cn c bnh thiu ht enzym chuyn ho phenylalanin
v tyrosin. Mt trong cc v d quen thuc v d tt bm sinh c di truyn do yu t gen ln Mendel
n trn nhim sc th thng vi tn s cao (khong 1/10.000) l Phenyl ceton niu. Bnh ny l do
thiu enzym phenylalanine hydroxylase trong gan c th oxy ho phenylalanine thnh tyrosin (hnh
8.11). Hu qu trc tip l tch t phenylalanine trong mu v bi tit thi qu tng phn qua nc tiu
di dng cc cht chuyn ho bt thng nh acid phenyl pyruvic.
8.5.1. t bin sinh dng vi sinh vt
Nm 1945, cc t bin lin quan n sinh dng c xc nh u tin nm Neurospora, sau

l vi khun. Nhiu vi sinh vt bnh thng s pht trin c trn mi trng ti thiu c cha ng,
ion NH4+ v mt vi ion cn thit khc. Nhng vi sinh vt ny l nhng vi sinh vt hoang di v c
gi l th nguyn dng. Sau khi c x l vi nhng tc nhn gy t bin cm ng, chng thu c
khng cn pht trin trn mi trng gin n na tr khi thm vo mt cht khc, thng l acid
amin, vitamin nucleotid hoc mt phn t vi lng no . Nhng c th mang t bin ny c gi l
th khuyt dng.
Nh vy, th nguyn dng c th tng hp tt c cc thnh phn t bo phc tp t cc nguyn liu

th s, n gin, nhng vi sinh vt khuyt dng th mt i kh nng ny. V h tng hp cc phn t
phc hp ni chung c lin quan n enzym v s t bin sinh vt khuyt dng c th do lm mt
hot tnh enzym.
8.5.2. t bin v bnh ngi
Vi nm sau pht minh ca Garrod, hng ngn bnh c tm ra c th di truyn c do t bin

gen. Nhiu bnh c lin quan n cc enzym b bt hot hay protein b thay i chc nng (bng 8.3).
Bng 8.3. Mt s bnh di truyn ngi c lin quan n protein chuyn bit
hay khim khuyt enzym
Tn s tnh
Triu chng Hot tnh sinh ho b nh
Loi ri lon Tn trn 1000
in hnh hng hay khim khuyt
ngi
Hng cu li Hng cu bin Hemoglobin c xu

10 - 20 (Afro -
lim (sickle cell dng, hnh li hng ngng kt v kt
Caribean)
anaemia) lim ta
Bnh thiu Thiu mt trong cc

mu a Trung Thiu mu chui polypeptid ca 10 - 20 (dn
Hi nng, lch ln hemoglobin ngi a Trung Hi)
(Thalassaemia) trng thnh
Mu
Thiu mu
huyt gii hng Thoi ho t
C nhiu dng tt c
cu bia (Non - bo hng cu, 0,5 (Pyruvat
u mt 1 enzym trong
spherocytic vng da, lch kinase)
nhnh chuyn ho HMP
hemolytic ln
anaemia)
Bnh a chy Mu khng ng Yu t VIII trong huyt

0,1 (n ng)
mu A c tng
Dystrophin, mt protein
Teo c
C Thoi ho c lin quan n mng t 0,1 (n ng)
Duchenne
bo c
Bnh Von ng huyt Glucose - 6 -

0,01
Gierke thp, sng gan phosphatase trong gan
Chuyn ho
carbohydrate
Khng dung y hi, tiu - Galactosidase trong
100
np lactose chy rut
Mc
phenylalanine,
Chuyn ho acid Phenylketon mc Phenylalanine
0,1
amin niu phenylacetic hydroxylase trong gan
acid trong huyt
tng v nc
tiu cao; tr tr
chm pht trin
Tn thng
Hypoxanthine guanin
Hi chng nron thn kinh,
Chuyn ho purin phosphoribosyl 0,1 (n ng)
Lesh - Nyhan thng c hnh
transferase
vi t hu hoi
Da nhy cm
vi nh sng,
gan b tn Enzym Ferro - chelatase
Chuyn ho Hem Protoporphyria thng, ca qu trnh sinh tng Him
protoporphyrin hp Hem
trong huyt
tng
Cc thc th lipoprotein
X va ng
t trng thp (LDL) trong
Vn chuyn Cao cholesterol mch
mng t bo LDL v 1
lipoprotein mu gia nh (Atherosclero -
cholesterol khng c
sis)
hp thu
Chm tr tu, tn
Thoi ho Bnh Acetylglucosamindase
thng gan v 0,01
Mucopolysaccharid Sanfilippo B trong lysosom
khp xng
B thoi ho
thn kinh, tch 0,3
Thoi ho BnhTay - Hexoseaminidase A
ly gangliosid (Ashkenazi
Gluosephingolipid Sachs trong lysosom
trong t bo Jews)
thn kinh
8.6. T BIN GEN V UNG TH
Rt nhiu nguyn nhn gy ung th khc nhau, nhng c ch phn t u lin quan n cc bin
i di truyn trn ADN lm tn thng tin trnh tng sinh bnh thng.
8.6.1. Cc nhm gen lin quan ti ung th
Lin quan ti s tng sinh ca t bo, c 2 con ng dn ti ung th:
- Th nht, cc bin i lm tng hot gen kch thch, kiu t bin ny c hiu qu tri. Alen bin
i c gi l oncogen (gen ung th), cn alen bnh thng l proto - oncogen (gen tin ung th).
- Th hai, cc bin i lm bt hot gen km hm, kiu t bin ny thng c hiu qu ln. Gen
km hm c gi l gen c ch khi u.
8.6.2. Gen ung th v gen tin ung th
8.6.2.1. Cc gen ung th do retrovirus ti np
Cc retrovirus bng con ung sao chp ngc tch hp c ADN ca mnh vo nhim sc th
t bo ch. Chng c th ngu nhin mang cc gen ung th lm bin i t bo ch bng cch xen on
ADN ca mnh cnh gen tin ung th ca t bo ch. Hng lot cc gen ung th c xc nh t
retrovirus gy bin i t bo (bng 8.4).
Bng 8.4. Mt s oncogen c xc nh nh retrovirus gy bin th
H gen Ngun gc
Chc nng ca gen tin ung th U do virus gy ra
ung th virus
abl Protein kinase (tyrosin) Chut, mo Pre - B - celleukemia
erb - B Protein kinase (tyrosin): th th ca G Erythroleukemia

yu t tng trng thng b (EGF)
Fibrosarcoma
Sis Yu t tng trng t tiu cu (PDGF) Kh Sarcoma, Carcinoma,

Myelocytoma
Src Protein kinase G Sarcoma
abl = Bch huyt Abelson; erb = Erythroblastose (tng nguyn hng cu);
Sis = Simian sarcoma virus; Src = Sarcoma.
Mt s gen ung th khc c pht hin khng do virus. Hin nay, pht hin c hn 60 gen
tin ung th.
Cc gen tin ung th hu nh lin quan n cc h thng tn hiu ca t bo nh cc protein, cc th

th xuyn mng, cc gen iu ho...
8.6.2.2. Cc cch bin i gen tin ung th thnh gen ung th
Ba cch ch yu trong cc cch l:
- Mt on hoc t bin im trong trnh t m ho.
- Khuch i gen.
- Cu trc nhim sc th.
Cc gen c th b bin i do t bin im, mt on, chuyn on nhim sc th hay do xen on

b gen ca retrovirus (hnh 8.12).
Hnh 8.12. Ba cch bin i gen tin ung th thnh gen ung th
8.6.3. Cc t bin gen c ch khi u
Qu trnh c ch l kt qu ca t bin th cp o nghch tc ng ca t bin s cp. Qu trnh

ny xy ra mc phin m v ch c hiu qu mt phn, lu tr protein c hot tnh cho t bo
sng m thng l mt loi gi hoang di bt thng.
Cc t bin gen c ch khi u thng l ln, nn t bo ch mt kim sot khi c 2 allel u b t

bin. C 2 t bin c pht hin:
- Gen Rb to protein retinoblasma c ch s pht trin mt dng ung th mt (retinoblastoma). Rb

cn c tc dng c ch i vi mt s dng ung th khc.
- Gen p53 to protein p53 c ch nhiu dng ung th. Cc nghin cu gn y cho thy protein p53
tham gia h thng cp cu, sa cha nhiu tn thng ca t bo ang phn chia.
Cc virus ADN nh Papilomavirus v SV40 c th gy ung th bng cch c lp cc sn phm ca

gen c ch khi u nh protein etinoblastoma hay protein p53.
8.7. CC H THNG CHN LC T BIN VI SINH VT
Cc h thng chn lc t bin gip pht hin c hiu qu cc t bin him hoi trong ngun t
bin rng ln. C nhiu h thng chn lc t bin v p dng tu thuc vo cc t bin (bng 8.5).
Bng 8.5. C s pht hin ba loi t bin da vo kiu hnh
H thng chn lc t bin c nh gi bng nng lc phn gii, l kh nng pht hin cc t
bin rt him so vi khng t bin. Nng lc phn gii cng ln th pht hin c cc t bin cng
him. V d, pht hin cc t bin khng c phn gii cao v khi cy s lng rt ln t bo ln
mi trng chn lc c cha cc tc nhn th phn ln t bo cht, ch c rt t t bo c t bin
khng mc c thnh khun lc.
8.7.1. Phng php khng
Dng tc nhn l thuc hoc phage chn lc vi khun. Cc t bin d dng c pht hin cc
khun lc mc trn mi trng thch c thuc hay phage.
8.7.2. Phng php lm giu chm
Dng pht hin c cc t bin khuyt dng. Dung dch vi khun pha long c cy trn b
mt mi trng thch ti thiu mc thnh cc khun lc ri. Ph mt lp mng mi trng tng t
ln mt mi trng thch va ri, cc khun lc bnh thng mc ln. Sau li ph thm mt lp
mi trng dinh dng c b sung v tip cho cht b sung khuch tn. Cc t bin khuyt dng s
mc sau khi c cht b sung, nn khun lc nh hn do mc chm.
8.7.3. Phng php lm giu hn ch
y l dng n gin hn ca phng php lm giu chm. Cc vi khun c cy trn mi trng

ti thiu c mt t cht b sung. Trong iu kin ny, cc t bin khuyt dng mc n khi ht cht b
sung th dng, do to khun lc nh. Cc vi khun bnh thng tip tc to khun lc to.
8.7.4. Phng php lm giu nh penicillin
Penicillin c tc ng dit khun dng phn chia. Vi khun c cho vo mi trng ti thiu c
penicillin. Cc vi khun ang tng trng b dit, ch cc t bo t bin khng tng trng cn sng
st. Sau , hn hp c cy ln mi trng khng c penicllin th cc t bin khuyt dng mc ln
vi t l tng i cao hn.
8.7.5. Phng php lc
Dng chn la cc t bin khuyt dng nm si. Dung dch bo t c nui trong mi
trng dinh dng thiu cht b sung. Cc t bin thiu cht b sung khng mc c, cc dng bnh
thng mc ra thnh nhiu si. Khi lc qua mng lc si thu tinh, cc dng bnh thng nhiu si b
gi li, cc dng t bin i qua mng lc. Dung dch c nhiu dng t bin c cy trn mi trng
c cht b sung v kim tra tm cc dng t bin.
8.7.6. Phng php in
Vi khun c cy mc ri tng khun lc trn mi trng dinh dng. Dng ming nhung (c
nhiu lng mn bc trn khc g) in ng v tr khun lc trn mi trng ti thiu. Cc khun lc t
bin khng mc ln c. Cn c v tr khun lc khng mc bn sao, tch cc t bin khuyt dng
(hnh 8.13).
Hnh 8.13. Phng php in

(a) a gc trn mi trng y ; (b) Khc g bc ming nhung;
(c) Mi trng c b sung cht A v B; (d) Mi trng c b sung cht A, B v C;
(e) Mi trng c b sung cht A v C
Tm tt
t bin l mt qu trnh c th c nghin cu mc sinh ho t bo v mc c th sinh

vt. t bin l nguyn nhn u tin ca cc bin d trong t nhin v trong th nghim, bin d cng c
th xy ra bng cch s dng cc ho cht v tia phng x gy t bin.
mc phn t, mt t bin l mt s thay i trong trnh t base ca ADN do mt sai st trong

s sao chp hay sa cha. t bin t nhin v tc ng ca cc tc nhn gy t bin c th c quy
v s bt cp sai ca cc base, trong khi cc li do cc h thng sa cha li t do thng thng li
tng ng vi s bin dng trong chui xon kp. S h hi ln ADN do tia cc tm c th hot ho
mt h thng sa cha li pht sinh.
Cc bin d t bin mc ADN c th l s thay th mt base ring l, hay chn hay xo mt

base (thay i khung) hoc ca vi base (a im). S thay th mt base ring l c th dn n s thay
i ca mt acid amin trong phn t protein to thnh v tu vo bn cht v v tr ca cc bin i

ny m protein c th mt hot tnh hon ton, hay mt hot tnh mt phn, hay c hot tnh nhng
khng n nh. Nu s bin i mt base lai to ra codon kt thc th protein s khng c tng hp
na v thng l bt hot. Cc t bin c ch cng c th bo ton c hot tnh trong mt s trng
hp nh s b tr cho t bin gc trong qu trnh dch m.
mc c th, cc kt qu t bin th hin mt tnh trng t bin no . Hu ht cc t

bin u c hi v cc t bin ny hu nh c bit cc bnh di truyn ngi. Hy vng gii thch
sinh ho s lm sng t cc nguyn nhn gy ung th.
Cc h thng chn lc t bin vi sinh vt hon thin cc phng php chn lc t bin trong
phng th nghim.
CU HI
1. Kiu hnh no khng l hu qu ca t bin vi khun?
a) Bin i t nguyn dng sang khuyt dng v ngc li.
b) t bin thnh thng bin v ngc li.
c) Mt hoc to thnh phn cu trc ca t bo.
d) Nhy cm hay khng thuc.
e) Nhy cm hay khng phage.
2. Trong bao nhiu bn m sao ca gen c mt t bin?
a) 105 b) 106
c) 107 d) 108
e) 109
3. Nhm tc nhn gy t bin nguy hi nht:
a) Base ng ng b) Alkyl ho
c) Deamin ho d) c ch tng hp base nit
Chn vo ADN
4. C ch khng o nghch sai hng do t bin:
a) Quang phc hi b) Lm mt nhm alkyl
c) Ni li bng ligase d) Sa sai bng glycosylase
e) b v c
5. C ch sa cha t bin bng cch loi b sai hng trong and:
a) Quang phc hi b) Lm mt nhm alkyl
c) Ni li bng ligase d) Sa sai bng glycosylase
e) Ti t hp
6. im khc bit gia k thut ti t hp v k thut t bin:
a) Ti t hp l k thut ghp gen
b) Ti t hp ph thuc vo k thut di truyn
c) t bin khng ph thuc vo k thut di truyn
d) a v b
e) a, b v c
7. Tn s t bin l mc xut hin ca t bin trn:
a) Mt nhim sc th b) Mt t bo
c) Mt giao t d) Mt ln sao chp
e) b, c, d
8. t bin t pht khng do
a) H bin ca base
b) Kh amin AP site
c) Kh amin to base ng ng
d) t bin lch khung khi polymerase sao chp cc on lp li ca mt nucleotid
e) B cm ng bi ho cht
9. C ch chng li t bin
a) NER - ct b nucleotid b) Mtase - kh alkyl
c) Glycosylase - ct b base sai d) Dung np AND sai
e) Tt c
10. Gen kch thch tr nn tng hot tnh gy ra bnh:
a) Ung th b) Tin ung th
c) Bnh thng d) Khng ung th
e) a v b
Bi 9
CC PHNG PHP PHN TCH ADN
MC TIU
- Nm c nguyn tc cc bc tin hnh chnh trong chit tch ADN.
- Nm c nguyn tc v ng dng ca k thut ct, ni, lai ADN.
- Trnh by c c s ca phng php ho hc v phng php enzym hc dng xc nh

trnh t acid nucleic.
- Nu c nguyn tc ca k thut PCR, cc bc c bn tin hnh v cc ng dng ca

n.
9.1. CHIT TCH ADN
ADN cha thng tin di truyn ca sinh vt, do mun nghin cu mc sinh hc phn t cn
chit tch ADN. ADN gm cc loi: ADN b gen, ADN ty th; ADN plasmid, ADN chloroplast v
ADN virus. Tu kch thc ADN, loi t bo m chn phng php chit khc nhau m bo chit
c ADN tinh khit v c hiu sut cao.
9.1.1. Phng php chit tch ADN
ADN l phn t c kch thc ln, khi chit tch cn lu trong thao tc trnh mi tc nhn c hc
hay ho hc qu mnh c th lm t gy phn t ny.
Phng php chit tch ADN gm ba bc chnh:
- Bc 1: Ph mng t bo v mng nhn (t bo nhn tht) bng cc tc nhn vt l hay ho hc

hoc kt hp nhiu tc nhn, gii phng ADN ra mi trng.
- Bc 2: Loi b cc thnh phn khng mong mun trong mu (protein, lipid, polysaccharid...).
Thng dng hn hp phenol - chloroform - iso amyl alchl (25 : 24 : 1) lm bin tnh protein, ng
thi khng ho tan acid nucleic; sau ly tm: protein bin tnh s ta thnh mt lp nm gia pha
nc cha acid nucleic v pha phenol - chloroform, thu nhn li pha nc cha acid nucleic.
- Bc 3: Kt ta acid nucleic. Thng dng cn ethanol th tch 2,5:1 v trong mi trng c lc

ion cao (nng mui cao) hoc dng isopropanol th tch 1:1. Cn acid nucleic thu c bng ly tm,
sau ho trong nc kh ion hoc dung dch m bo qun.
9.1.2. Tinh ch
9.1.2.1. Tinh ch acid nucleic bng siu ly tm
- Siu ly tm trn gradient lin tc cesium chlorid (CsCl). K thut ny c th t phn gii 1%
n v t trng, cho php phn tch hai phn t ging nhau trong mt phn t c nh du bng
mt ng v nng.
- Siu ly tm trn m cesium chlorid (gradient khng lin tc). K thut ny thng c p dng
tinh ch mt lng ln acid nucleic.
- Siu ly tm trn gradient saccharose. K thut ny dng phn tch hn hp cc acid nucleic c
kch thc chnh lch nhau nhiu kb.
9.1.2.2. Tinh ch acid nucleic bng sc k
- Sc k i lc: s dng pha tnh l U - Sepharose hay oligodT - cellulose tinh ch mARN.
- Sc k lc gel tch cc nucleotid t do sau khi to mu d nh du.
- Sc k trao i ion trn vi ct, p dng thu hi nhng lng ADN rt nh.
- Sc k lng hiu nng cao: dng tinh ch cc oligonucletid tng hp ( phn gii l 1
nucleotid), plasmid, phn tch cc on ADN.
9.2. CC PHNG PHP PHN TCH NH TNH V NH LNG S B ACID

NUCLEIC
9.2.1. Quang ph k
Phng php ny cho bit kt qu tng i nng acid nucleic trong mu.
Nguyn tc phng php da vo s hp th mnh nh sng t ngoi bc sng 260 nm ca cc

base purin v pyrimidin. Gi tr mt quang bc sng 260 nm (OD260nm, Optical Density bc
sng 260 nm) cho php xc nh nng acid nucleic trong mu da vo tng quan sau:
Mt n v OD260nm tng ng vi mt nng l:
50 g/ml dung dch ADN (hoc ARN) si i
40 g/ml dung dch ARN (hoc ADN) si n
30 g/ml oligonucleotid (ti 70 base)
V d: gi tr OD260nm = 0,9 s tng ng vi:
Dung dch c nng ADN si i = 0,9 50 = 45 g/ml
Dung dch c nng ARN si n = 36 g/ml
Tuy nhin cch tnh ny ch ng vi cc dung dch acid nucleic tinh khit. kim tra tinh khit
ca dung dch, ngi ta o thm gi tr OD 280 nm (OD280nm). 280 nm l bc sng cc protein
c mc hp th cao nht, nhng cc protein cng hp th nh sng bc sng 260 nm nh cc acid
nucleic nn lm sai lch gi tr tht ca nng acid nucleic. Mt dung dch acid nucleic c xem l
tinh khit (khng tp nhim protein) khi t s OD260nm/OD280nm nm trong khong 1,8 - 2.
9.2.2. in di phn tch ADN
Phng php ny c p dng c trong phn tch nh tnh ln trong vic thu nhn mu acid
nucleic.
Nguyn tc phng php da vo c tnh cu trc ca cc acid nucleic. l cc i phn t tch

in m ng u trn khp b mt nn khi chu tc ng ca mt in trng, chng s di chuyn v
hng cc dng. S di chuyn nhanh hay chm tu thuc vo hnh dng v t s in tch: khi lng,
nh cc cht khc nhau s c tch ra thnh tng vt ring v c pht hin bng nhiu cch khc
nhau. Tuy nhin, hu ht cc phn t ADN u c hnh dng v t s in tch - khi lng nh nhau do
khng tch c bng in di thng thng, m phi dng k thut in di trn gel. Gel c lm
bng agarose hay polyacrylamid c cc l do cc dy polyme agarose hay polyacrylamid an vi nhau
thnh mng li to ra, phn t ADN khi phi chui qua cc l ny n in cc, do phn t
ADN nh s di chuyn nhanh hn phn t ln, nh ta tch c ADN theo kch thc phn t (c
ch ry).
Tc in di ca ADN qua gel tu thuc vo bn thng s chnh: (1) Kch thc phn t ADN - l
thng s quyt nh nht. Phn t ADN si i dui thng di chuyn vi tc t l nghch vi logarit
ca trng lng phn t ca chng; (2) Cu dng ca ADN, 3 dng tn ti ca mt ADN l dng siu
xon, dng vng v dng thng tuy c cng trng lng phn t v thnh phn, nhng di chuyn qua gel
vi tc khc nhau; (3) Nng gel xc nh tc di chuyn v hiu qu tch cc phn t ADN, s
di chuyn ca phn t ADN t l nghch vi nng ca gel; v (4) in th s dng - in th thp,
tc di chuyn t l vi in th s dng.
in di trong trng xung c p dng phn tch cc phn t ADN rt ln (>50 kb). Nguyn
tc phng php da vo s i hng in trng trong qu trnh in di. Mi ln hng in trng
thay i th phn t ADN phi t nh hng li. Thi gian cn cho vic t nh hng li tu thuc
kch thc ca phn t, phn t cng di th thi gian nh hng li cng ln khin cho n di chuyn
chm hn phn t kch thc nh.
i vi ADN b gen nhim sc th ca ng vt c v c kch thc ln (khong 109 kDa), khng

th phn gii bng in di agarose thng quy, ngay c bng in di trong trng xung, phi thu phn
ADN vi enzym gii hn trc khi phn tch.
9.3. K THUT CT, NI, LAI ADN V NG DNG
9.3.1. Ct ADN bng enzym ct gii hn
9.3.1.1. Khi nim v enzym ct gii hn
T bo vi khun c mt h thng bo v chng li s tn cng ca phage l cc enzym ct gii

hn (RE). H thng ny gm hai loi enzym: (1) Cc RE ct ADN phage nhng v tr chuyn bit v
(2) Cc enzym methylase gn nhm methyl vo A hay C trn ADN t bo vi khun v tr ct ca cc
RE nn ADN ca vi khun khng b chnh cc RE ca chng ct.
Da vo kh nng nhn bit v ct mt trnh t xc nh trn phn t ADN, ngi ta chia RE thnh
ba loi: (1) Loi I - enzym ct phn t ADN ti im cch trnh t nhn din khong 1000 - 5000
nucleotid v gii phng vi chc nucleotid; (2) Loi II enzym ct ngay v tr nhn bit trnh t; (3)
Loi III - enzym ct phn t ADN ti im cch trnh t nhn din khong 20 nucleotid. Trong k thut
di truyn, cc RE loi II c s dng rt nhiu thao tc trn ADN nn sau y ch trnh by cc c
tnh ca loi ny.
9.3.1.2. c im ca RE loi II
Trnh t c trng RE nhn bit thng bao gm 4 - 8 nucleotid. Mi enzym ch nhn nhn mt
trt t chuyn bit c cu trc palindrome, ngha l hai mch ca trnh t hon ton ging nhau khi
chng c c theo chiu 5 3, v d trnh t GAATTC ca enzym EcoRI.
Cc enzym ct ADN gm hai loi: exonuclease v endonuclease. Exonuclease ct ADN t hai u

mt, cn endonuclease ct ADN gia phn t. Cc RE ct ADN gia mt cch c hiu, c gi l
endonuclease gii hn.
T nm 1962, khi Werner Arber ln u tin pht hin ra RE trong t bo vi khun, ti nay ngi ta
pht hin hn 500 loi RE v hn 100 loi ang c thng mi ho.
Cc RE loi II c hai kiu ct: (1) ct to u t do RE ct hai mch ADN ti cng mt im v (2)
ct to u so le hay u dnh. Cc enzym ct to u dnh c s dng rt nhiu trong ti t hp di
truyn in vitro, hai phn t ADN c ngun gc khc nhau nhng cng c ct bi mt RE s c kh
nng kt hp thnh mt thng qua cc u dnh.
9.3.1.3. ng dng
Vic s dng cc RE c ngha quan trng trong nghin cu di truyn phn t. Chng cho php ct
nh b gen khng l ca cc sinh vt t bo nhn tht, lp bn gii hn, so snh b gen cc loi
khc nhau thng qua k thut RFLP.
9.3.2. Cc ligase v ng dng ni cc on acid nucleic
Cc enzym ligase xc tc s hnh thnh lin kt ni hai on ADN (ADN ligase) hay ARN (ARN
ligase).
Cc enzym ny c th c ly trch t E. coli (E. coli ADN ligase) hoc t phage T4 xm nhim E.
coli (T4 ADN ligase v T4 ARN ligase).
E. coli ADN ligase c kh nng xc tc phn ng ni hai trnh t ADN c u dnh; T4 ADN ligase
cng chc nng vi E. coli ADN ligase nhng c bit cn c kh nng ni hai trnh t ADN c u
bng; v T4 ARN ligase xc tc ni hai trnh t ARN bng lin kt phosphodiester thng dng
nh du phng x u 3 ca cc phn t ARN, sn xut on d.
9.3.3. K thut lai nucleotid v ng dng
9.3.3.1. Khi nim v lai phn t
Khi nhit c nng ln ti nhit "bin tnh" (Tm), hai mch phn t ADN tch ri nhau, s
bt cp tr li s khng xy ra nu nhit phn ng h xung t ngt v phn t ADN s tn ti trong
mi trng dng mch n di mt cu hnh khng gian v trt t. Ngc li, nu sau khi hai mch
tch ri, nhit c lm gim t t cng vi iu kin thch hp, hai mch s bt cp tr li. Hin
tng ny c gi l s lai phn t, n c tnh c hiu tuyt i.
Cc yu t nh hng n s lai phn t:
- Nng ADN v thi gian phn ng.
- Nhit .
- di ca cc trnh t.
- Lc ion.
Cc phng php lai phn t rt a dng, trong bi ny ch cp kiu lai trn pha rn v lai ti ch.
9.3.3.2. Lai trn pha rn v ng dng
Nguyn tc ny l mt trong hai trnh t b sung (cc mch n) nm trong mi trng lng l mt
dung dch m v trnh t kia (thng l trnh t ch, tc trnh t cn tm) c c nh trn mt gi th
rn.
Cc phng php lai trn pha rn c ng dng nhiu nht l Southern blot, Northern blot v dot
blot.
Phng php Southern blot do Southern m t nm 1975, c s dng nh v nhng trnh t

c bit trn ADN b gen, hay nhng ADN kch thc nh nh ADN plasmid, phage,... Phng php
ny c p dng lp bn gii hn ca mt gen; pht hin cc a dng trnh t ca cng mt
gen cc chng hay cc c th khc nhau qua s so snh bn gii hn ca chng; pht hin cc t
bin mt on, t bin im hay ti t hp trn mt gen v chng lm thay i bn gii hn.
Hnh 9.1. S phng php Southern blot
Phng php Northern blot c s dng xc nh kch thc v hm lng ca mt mARN c

trng trong mt hn hp ARN.
Phng php dot blot cho php xc nh tng i mt ARN c trng trong mt hn hp ARN m
khng cn phi phn tch chng ra. Phng php ny c th s dng cho ADN.
9.3.3.3. Lai ti ch
Nguyn tc lai ti ch l mt kiu lai phn t trong trnh t acid nucleic cn tm (trnh t ch)
nm ngay trong t bo hay trong m. Lai ti ch cho php nghin cu acid nucleic m khng cn phi
tch chit chng ra khi m, t bo.
Cc ng dng ca kiu lai ny rt a dng i t k thut nh v gen trn nhim sc th, pht hin
dng vi khun ti t hp trong phng php to dng n vic nghin cu mt mARN chuyn bit trong
t bo v m.
9.3.3.4. Cc loi mu d
Mu d l bn sao ca mt trnh t acid nucleic v c nh du d nhn bit. Mu d c tnh

(1) chuyn bit do ch lai vi ng trnh t bt cp b sung v (2) nhy do c nh du pht hin
v nh lng. Tc nhn dng nh du mu d l cc ng v phng x: 32P, 35S, 3H, hoc ho cht:
h thng avidin (hay streptavidin) - biotin, h thng peroxydase - c cht.
9.4. CC PHNG PHP XC NH TRNH T ADN
9.4.1. Phng php Maxam v Gilbert - phng php ho hc
Phng php gm cc bc chnh nh sau:
- nh du cc phn t ADN bng 32P mt u.
Hnh 9.2. S phng php Maxam v Gilbert
- Chia hn hp phn ng thnh nm phn on, mi phn on chu mt x l ho hc chuyn bit

lm bin i c trng mt loi nucleotid v sau ct phn t ADN ngay nucleotid . Nm phn on
tng ng nm nhm nucleotid b tc ng l: G, A, C, G + A, T + C. Sau khi x l, mi phn on l
mt tp hp oligonucleotid c kch thc khc nhau nhng u chm dt ti cng mt loi nucleotid.
- Phn tch trn gel polyacrylamid, v tr ca cc oligonucleotid trn gel tng ng vi kch thc
ca chng v c pht hin nh u c nh du phng x.
9.4.2. Phng php Sanger v cng s phng php enzym
- To dng trnh t ADN cn xc nh trong mt vect mch n (phage 13).
- Tng hp ADN trn khun ADN mch n nh enzym ADN polymerase, vi s hin din ca bn
loi nucleotid, trong c mt loi c nh du ng v phng x (35S).
Phn ng tng hp c tin hnh trong bn phn on ring, mi phn on c cho mt trong
bn loi dideoxyribonucleosid (l nhng deoxyribonucleosid c nhm 3OH c thay bng H) vi hm
lng rt nh. Do hm lng thp nn thnh thong mi c mt dideoxynucleotid c s dng vo
phn ng tng hp mt oligonucleotid, v lp tc s tng hp oligonucleotid dng li.
- Phn tch trn gel polyacrylamid v kt qu c c trn bn phng x t ghi.
Hnh 9.3. Kt qu nh trnh t theo phng php Sanger
C mt s phng php khc c ci bin t phng php Sanger, n gin hn v rt gn hn,
l phng php xc nh trnh t bng my t ng v phng php PCR dng trong xc nh trnh
t acid nucleic.
Hnh 9.4. Kt qu nh trnh t bng my nh trnh t t ng
9.5. PCR
9.5.1. M u
PCR l phn ng tng hp ADN nh enzym polymerase do Karl Mullis v cng s pht minh nm
1985. PCR a li mt cuc cch mng trong di truyn phn t: khuch i in vitro acid nucleic c
chn lc bng to dng in vitro m khng cn s hin din ca t bo, khc phc c nhc im thao
tc phc tp v cn thi gian di ca k thut to dng in vivo. Nh PCR, mt on ADN mt vng
bt k trong b gen c khuch i ln rt nhiu ln khi trnh t nucleotid hai u on bit.
9.5.2. Nguyn tc c bn
PCR s dng cc c im ca qu trnh sao chp ADN. ADN polymerase dng cc on ADN
mch n lm khun tng hp nn si mi b sung vi si ny. Cc si ADN khun mch n c
th to ra theo cch n gin l un nng dung dch ADN mch kp n gn nhit si.
Tt c cc ADN polymerase khi hot ng tng hp mt mch ADN mi t mch khun mu u

cn s hin din ca nhng mi chuyn bit. Mi l nhng on ADN ngn, c kh nng bt cp b
sung vi mt u ca mch khun, v ADN polymerase s ni di mi hnh thnh mch mi. Phn
ng chui c hnh thnh da vo c tnh ca cc ADN polymerase. Tht vy, nu ta cung

cp hai mi chuyn bit bt cp b sung vi hai u ca mt trnh t ADN, ta s ch tng hp on
ADN nm gia hai mi. iu c ngha l khuch i mt trnh t ADN xc nh, ta phi c thng
tin ti thiu v trnh t to cc mi b sung chuyn bit; cc mi ny gm mt mi xui v mt
mi ngc (so vi chiu phin m ca gen).
Hnh 9.5. Phn ng chui polymerase
Phn ng chui l mt s khuch i in vitro mt trnh t ADN khun mu vi xc tc enzym. Phn

ng ny l s ko di ng thi on mi ca cc mch b sung ADN. Bng cch lp li cc chu k
gm vic a nhit ln 95oC (khong 35 chu k), h nhit xung (37oC 65oC) v ko di
72oC bng cch s dng mt loi ADN polymerase chu nhit. Phn ng c thc hin trong mt ng
nha t trong mt my c th iu chnh nhit gi l my chu k nhit (my PCR), my ny c th
ci chng trnh lm nng v lm ngui theo chu k cn thit. Qu trnh phn ng s lm gia tng s
bn sao mt trnh t khun mu ban u (hnh 9.5).
C hai si ADN u c dng lm khun cho qu trnh tng hp nu cc on mi c cung

cp cho c hai si. Trong phn ng PCR, cc on mi c chn chn hai u ca on ADN cn
nhn ln, sao cho cc si ADN tng hp mi c bt u t mi on mi v ko di v pha on mi
nm trn si kia (hnh 9.6c). Nh vy, sau mi chu k cc im bm cho cc on mi li xut hin trn
mi si ADN mi c tng hp.
Hn hp phn ng li c un nng ln tch si ban u khi si mi tng hp, cc si ny sau
li c dng tip cho chu trnh tip theo, bao gm cc bc gn on mi, tng hp ADN v
tch ri cc on. Kt qu cui cng ca phn ng PCR l sau n chu k phn ng, tnh theo l thuyt ta
s c 2n-2 bn sao cc phn t ADN mch kp nm gia hai on mi (bng 9.1).
Bng 9.1. Hiu qu nhn ADN ca PCR
S phn t ADN mch S phn t ADN mch

S chu k (n) S chu k (n)
kp sinh ra kp sinh ra
1 0 17 32768
2 0 18 65536
3 2 19 131072
4 4 20 262144
5 8 21 524288
6 16 22 1048567
7 32 23 2097152
8 64 24 4194304
9 128 25 8388608
10 256 26 16777216
11 512 27 33544432
12 1,024 28 67108864
13 2,048 29 134217728
14 4,096 30 268435456
15 8,192 31 536870912
16 19,384 32 1073741824
Hnh 9.6. S nguyn l phn ng chui polymerase

n gin ho s , t phn (d) khng v hai si ADN khun ban u
9.5.3. Cch tin hnh mt phn ng PCR
PCR l mt chui cc phn ng gm nhiu chu k ni tip nhau. Mi chu k gm ba bc:
Bc 1: Trong mt dung dch phn ng bao gm cc thnh phn cn thit cho s sao chp, phn t
ADN c bin tnh nhit cao hn nhit "chy" Tm ca phn t, thng l 94oC 95oC trong
vng 30 giy 1 pht. Ti nhit ny, cc phn t ADN mch kp tch nhau ra hon ton, to nn cc
si n dng lm khun cho cc on mi v ADN polymerase. y l giai on bin tnh.
Bc 2: Nhit c h thp (thp hn Tm ca cc mi), cho php cc on mi bt cp vi

khun. Trong thc nghim nhit ny dao ng trong khong 40oC 70oC, tu thuc vo Tm ca cc
mi s dng v ko di t 30 giy n 1 pht. y l giai on lai.
Bc 3: Nhit c tng ln n 72oC gip cho ADN polymerase chu nhit hot ng tt nht.
Thi gian tu thuc di ca trnh t ADN cn khuch i, thng ko di khong 30 giy hay hn
na. on ADN nm gia hai mi c tng hp. y l giai on tng hp hay ko di.
Vo cui giai on ny, nhit mt ln na c nng ln 94oC, nhng ln ny ch trong vng 30

giy cho cc mu ngn ca ADN mch kp (c si ban u v si mi c tng hp) tch nhau ra.
Cc si n ny tr thnh khun cho mt chu k tng hp ADN khc.
Tm li, mt chu k gm ba bc: bin tnh (un nng) tch cc si ADN kp thnh si n
lm khun, lai ho (gn kt) cc on mi vo si khun v ko di (tng hp) bng ADN polymerase,
s c lp i lp li nhiu ln v mi ln li lm tng gp i lng mu ca ln trc. y l s
khuch i theo cp s nhn (hnh 9.6).
9.5.4. Cc thnh phn phn ng
Vt liu gm trc ht l ADN c cha on cn nhn ln. Khng cn thit phi tch ring on cn
nhn ln v vic c cc on mi dng trong phn ng xc nh. Hm lng ADN cn cho phn
ng rt nh. Trong cc th nghim bnh thng phng th nghim ch cn mt microgram ADN b gen
l . Trong nhiu trng hp, phn ng PCR c th nhn cc on ADN ch t mt phn t ring l.
Hai on mi xc nh im bt u tng hp ADN, ADN polymerase, v hn hp ca 4
deoxyribonucleotid ( dng dNTP) v dung dch m c ion Mg2+ cn c a vo ng nghim c
cha ADN khun. Dung tch tng s thng l 50 l hoc 20 l. Hn hp phn ng c cho vo ng
nha nh chu nhit, c np y.
Mt phn ng PCR c th gm cc thnh phn hay cc vt liu ban u: mch khun cho s khuch
i, cc on mi oligonucleotide, dNTP, v ADN polymerase chu nhit vi dung dch m phn ng
thch hp v MgCl2. Nng ca mi thnh phn ny tu yu cu. Tr ADN khun v cc on mi,
cc thnh phn cho PCR u c sn t cc nh cung cp.
9.5.4.1. Cc polymerase chu nhit
Enzym ADN polymerase c s dng u tin trong PCR l on Klenow ca ADN polymerase I.
c tnh ca on ny l xc tc s tng hp ADN theo chiu 5 - 3 v c hot tnh exonuclease (thu
gii lin kt gia cc nucleotide t u phn t ADN) theo chiu 3 - 5. Tuy nhin enzym ny
khng chu c nhit nn thao tc phc tp v hiu qu thp (phi thm enzym mi vo phn ng sau
mi ln bin tnh v enzym c b nhit phn hu, nhit lai thp khin s khuch i khng c hiu
vv...).
Hin nay thng dng ADN polymerase l Taq polymerase chu nhit, c tch chit t mt loi vi
khun sui nc nng c tn Thermus aquaticus. Enzym ny khng b ph hu nhit bin tnh v
xc tc s tng hp t u n cui qu trnh phn ng.
Taq polymerase c sn trn th trng t cc cng ty cng ngh sinh hc. Cc polymerase chu nhit
khc c trn th trng bao gm Vent polymerase c phn lp t Thermococcus litoralis, xc tc tng
hp nhiu bn sao chnh xc t mch khun hn l Taq polymerase. Ngoi ra cn c Tth polymerase
c phn lp t Thermus thermophilus. Cc polymerase chu nhit l thnh phn t nht ca PCR v
thng c s dng 1 n v cho mt phn ng. Hm lng ny xc tc cho s khuch i
khong 40 chu k bng my PCR.
Trc y, vic la chn polymerase chu nhit cho 1 th nghim th khng my kh khn v ch c
mt loi ADN polymerase chu nhit (Taq). Hin nay c nhiu loi polymerase c ngun gc khc nhau,
nhiu bn sao ca cc polymerase ny c bin i di truyn v chng c th u vit hn polymerase
gc trong phm vi mt vi ng dng (bng 9.2). S chn la mt polymerase chu nhit cho mt th
nghim PCR ph thuc vo yu cu ca tng th nghim.
Bng 9.2. Cc loi ADN polymerase thng gp
Mc n nh Hot ng Cation cn 3 5
Enzym Ngun gc Ni 95oC (t1/2) ti u thit Exonuclease
Thermus Sui nc MgCl2
Taq 40 pht 50 - 75 -
aquaticus nng
AmpliTaq Tag ti t hp 40 pht 50 - 75 MgCl2 -
AmpliTaq AmpliTag ct
b 289 aa N - 80 pht 75 - 80 MgCl2 -
Stoffel tn
Thermus Sui nc MnCl2 (RT),

Tth 20 pht 55 - 75 -
thermophilus nng MgCl2 (PCR)
Dng chy
Thermotoga
Ultima nng +
maritima
bin su
Dng chy
Thermococcus >95% hot MgSO4
Vent nng 80 +
litoralis ng sau 1 h
bin su
Vent (Exo - Vent ti t MgSO4 -
) hp
Pyrococcus Dng chy
>95% hot MgSO4
Deep Vent sp. Strain GB nng 72 - 80 +
ng sau 1 h
-D bin su
Dng chy
Pyrococcus >95% hot MgCl2
Pfu nng 75 +
furiosus ng sau 1 h
bin su
9.5.5.2. ADN khun mu
Phn ng chui cc nhy, c th ch cn dng mt phn t ADN lm khun cng thu c sn

phm. Tuy nhin, mc nhy cm cao l nguyn nhn chnh gy ra s dng tnh gi do cc ADN
ngoi nhim.
Phn ng khuch i ti u xy ra trn ADN tht tinh khit nhng nhiu k thut chn on bng
PCR vn t kt qu tt vi ADN thu nhn trc tip t dch chit mu th (vt mu, vt liu lu tr,
ADN c v ngay c vi khun tit trng). Tuy nhin, trong nhiu trng hp khc, cc qu trnh chun
b mu phi c thc hin m bo cho vic khuch i, v cc cht c ch nh heparin, EDTA, hay
cc acid c th hin din trong mu.
Mch ADN khun mu cng c th nh hng n phn ng chui. Nu ADN ny c cha nhiu
GC th s kh khn cho vic tch ri cc chui ADN nhit cao. Trong cc trng hp ny, vic
thm formamide hay dimethyl sulfoxide (DMSO) vo hn hp phn ng c th gii quyt c vn .
Nng ca ADN khun mu trong PCR s nh hng n mc khuch i. Vi vic s dng

cc polymerase cho hiu qu cao, lng ADN mu s dng cng c khuynh hng gim (1 microgram
cn 100 nanogram). Mc d, cc khun mu n c khuch i nhng khng phi mi th nghim
u t c s nhy cm ny. Do , nn thit lp PCR vi lng ADN khun mu thch hp. Lng
khun mu ADN d tha c th nh hng xu n qu trnh khuch i. Nu lng ADN khun mu
qu cao th s c khuynh hng to ra lng ln cc sn phm c ko di t on mi hn l nhng
sn phm nm gia cc on mi. Vic la chn mi, qu trnh lp li khng thch hp v s c mt
ca cc cht c ch cng c th lm gim s khuch i.
ADN khun c th l b gen tch ra t t bo, ADN t cc ngn hng genome hoc cADN (ch cn
nng di ng), cc ADN bt k, ARN ton b hoc ARNm.
9.5.4.3. Cc on mi
Cc on mi l thnh phn quyt nh s khuch i c trng v c hiu qu cao ca PCR. Vic

chn on mi phi tun th mt s nguyn tc:
- Trnh t ca mi c chn sao cho khng th c s bt cp b sung gia cc mi to cu trc bc

hai hay to dimer gia cc nucleotid ngay trong cng on mi lm cho phn ng gim hiu qu v
ngn cn s to thnh on mong mun.
- Cc mi chn phi c trng cho trnh t ADN cn khuch i, khng trng vi cc trnh t lp li
trn gen.
- Kch thc mt on mi ti u t 18 - 25 nucleotid. on mi vi kch thc ny m bo cho

vic gn c hiu nhit ti u v gi thnh cho mt on mi c gim thiu.
- Khong cch gia hai mi khng di qu 3 kb. Phn ng PCR s ti u trn nhng trnh t nh
hn 1 kb. Tuy nhin hin nay c mt s enzym polymerase chu nhit c th tng hp c on ADN
c kch thc khong 30kb.
- T l GC cc mi thng chim khong 40 - 75%. Trong thnh phn cc on mi trnh c qu

nhiu cp GC hay l mt lot cc nucleotid G v C. V lin kt G: C mnh hn lin kt A: T (G ni vi
C bng 3 lin kt hydro cn A vi T ch c 2). Do vy, vic b gy lin kt gia G: C cn nhiu nng
lng hn lin kt A: T. Ngi ta c th tnh c nhit ti u cho vic cc on mi trong

PCR, nhit ny gi l Tm 5oC l nhit "chy" ca on mi tr i 5. Kt qu ny c t php tnh
n gin:
Tm = 2(A + T) + 4(G + C)
Nhit = Tm 5oC
Cc on mi c tng hp ho hc trn mt cht ti rn trong my tng hp oligonucleotide. Cc

my ny c t ng ho cao, d dng tng hp ra cc on mi. C nhiu quy tc thit k on
mi v c mt s phn mm cho s chn la mt cp on mi ti u cho s khuch i ca mt
ADN khun mu c trnh t bit. Nng mi khong 0,1 M n 1 M. Nng thng c xc
nh da vo gi tr OD (optical density) o 260 nm.
9.5.4.4. Magnesium chloride
Nng MgCl2 nh hng n s thnh cng ca phn ng, nu khng c mt Mg+2 th s khuch
i s khng xy ra. MgCl2 cn thit cho hot ng ca ADN polymerase khi ADN vi cc on mi
v n c mt mi tng quan vi nucleotid triphosphat. Nng MgCl2 ti u cho PCR phi c xc
nh cho tng phn ng bng thc nghim. C th dng phng php nh phn bng cch thit lp mt
lot phn ng vi cc nng MgCl2 khc nhau t 1 mM n 5 mM v xc nh nng MgCl2 ti u
cho mu mong mun.
9.5.4.5. Nucleotid triphosphat (dNTP)
Bn loi nucleotide thng c s dng nng l 20 - 200 M/ mi loi nucleotid. Nng

cao hn d dn n s khuch i "k sinh". S mt cn bng trong thnh phn cc nucleotid lm tng
cc li sao chp ca polymerase.
9.5.5. Chu k nhit PCR
Phn ng chui khuch i ADN t c bng cc chu k c lp i lp li. Mi chu k gm s

bin tnh bng cch nng nhit , bng cch h nhit v s ko di. Mt chu k in hnh cho vic
khuch i on ADN gm 500 cp base s l bin tnh 94oC trong 60 giy, 50oC trong 60 giy v
72oC trong 60 giy. Mi PCR c thit k v c ti u ho khc nhau v tu thuc mt s cc yu
t:
- Kch thc ca on c khuch i. Cc sn phm khuch i ln hn c th cn giai on bin

tnh v ko di lu hn. Cc on ngn hn cn t thi gian hn giai on ko di.
- Trnh t ca cc on mi v cc iu kin v ion s quyt nh nhit tt nht cho mt chu k.

V vy, da vo cc phn ng c thc hin, nhit c th thay i t 37oC n 65oC.
9.5.6. u im v nhc im ca phng php PCR
Vic ng dng rng ri k thut PCR c nhiu l do:
- Rt nhanh chng. Ch tn khong 3 gi l c th khuch i mt trnh t mong mun bit so vi
cc k thut cng ngh di truyn "c in" phi mt mt tun hay hn na.
- n gin: PCR c th c thc hin trong mt ng nh vi cc thnh phn ti thiu v ch vic

trn chng li vi nhau. Cc phng php to dng gen in hnh khc cn c cc vt liu mc tin nh
cc mng v cc nucleotid triphosphate c nh du phng x v cc k thut c bit. PCR c th
c thc hin trn cc mu c cha ADN tng i th, v d vt mu cha c x l cho vic phn
tch php y. iu ny ngc vi cc phng php v thao tc gen l cn phi c ADN c khun mu ln
vector tng i tinh khit. Cc yu t trn cho thy rng PCR l mt s thay i ng k so vi cc
phng php c in cho vic khuch i cc trnh t c bit.
- Nhy: Phn ng PCR cc nhy v c th ch cn dng mt phn t ADN lm khun cng thu c
sn phm.
Tuy nhin, cn phi nhn mnh rng cc ng dng rng ri ca k thut PCR i hi phi bit r
trnh t ADN khun mu. y l mt im gii hn ca k thut ny, ngha l n vn phi da vo nn
tng ca cc k thut cng ngh di truyn ch khng hn l thay th cc k thut ny. Do vy ngi ta
khng s dng k thut ny trong cc k thut to dng gen thit k. Trong mt vi trng hp phng
php PCR khng p dng c vi cc on ADN kch thc ln hn 3 kb (thng 1 kb l tt nht).
Ngoi ra kh nng ngoi nhim i vi phng php ny l rt ln.
9.5.7. Cc phng php PCR ci tin
Phn ng PCR c p dng rng ri v c nhng bin i ph hp vi tng mc ch nghin cu

ring. V vy chng ta gp nhiu phng php khc nh PRC t (Nested - PCR), ADN nhnh (Branch -
ADN), PCR a thnh phn (multiplex - PCR), PCR ngc (Reverse - PCR), RAPD - PCR... nhng tt
c u da trn nguyn tc ca phn ng PCR.
9.5.7.1. PCR "t"
t c nhy cao trong PCR, ngi ta c th thc hin 2 PCR lin tip. Trn mt in di
ca cc on thu c sau PCR th nht, ngi ta quan st thy ngoi mt vt chnh cn c cc vt nh
khc. iu ny l do cc mi c th bt cp vi vng khc ca ADN ch bi s lai ho khng c hiu.
PCR th 2 c thc hin vi cc mi ging PCR 1, khuch i cc sn phm thu c t PCR th

nht. PCR "t" c c hiu cao do s dng 2 cp mi khc nhau.
Cp mi ngoi: Cp mi ny to ra cc on ADN c khuch i ging vi PCR c in. Chng

c hiu cho 2 trnh t mt ca ADN cn khuch i. Cc on ADN thu c lm khun mu cho PCR
th 2.
Cp mi ni: Cp mi ny b sung vi vng

nm bn trong chui nucleotid thu c vi cp
mi th nht. Thut ng "PCR t" l v cc on
mi c t, ng hp trong ln PCR u.
K thut ny c nhy v chuyn bit cao.

Ngi ta dng k thut ny pht hin virus gy
bnh vim gan C, mt loi virus c vt liu di
truyn l ARN. ARN virus khng th c pht
hin bng s lai ho trc tip, nn phi thc hin
phn ng chui khuch i vt liu di truyn

ca virus. PCR th nht chuyn tt c ARN virus
thnh cADN, y chnh l bc sao m ngc nh
enzym phin m ngc to cADN bng k thut
PCR "t". Trong PCR th 2, mt mi c biotin
ho, mt mi c gn vi iod 125. Cc on thu
c s c gn trn mt gi mang avidin (avidin
l mt cht c i lc mnh i vi biotin), v cui
cng ngi ta o hot tnh phng x kt lun
mu l dng tnh hay m tnh (hnh 9.7).
PCR "t" c nhy cao hn PCR tiu chun l

do lp li s khuch i sn phm t mt PCR th 1
vi mt PCR th 2. Tuy nhin vn ngoi nhim
cng l vn hn ch ca n.
Hnh 9.7. PCR "t" pht hin HCV
9.5.7.2. ADN "nhnh"
ADN ch c khuch i bng PCR, sau cho lai vi on d (l mt phn c hiu ca ADN
ny). Tn hiu c lai ho c hiu vi on d theo cng cch thc ca ADN ch c gn (cht hunh
quang, cht d quang hay cht mu), tn hiu s c to ra nh cht nh du hin din sau cng my
khuch i. PCR "nhnh" c c hiu cao.
K thut ADN "nhnh" c dng pht hin ADN ca virus gy bnh vim gan B. Trong k
thut ny s dng 5 on d. Mt on d thnh phn l mt trnh t cho php s lai ho cng mt lc
trn 2 on ch khc nhau.
Hnh 9.8. K thut ADN "nhnh" s dng 5 on d
9.5.7.3. PCR a thnh phn
Trong k thut ny, 2 hay nhiu ADN ch c khuch i ng thi. PCR a thnh phn c th
c s dng thng xuyn trong cc xt nghim chn on, s dng 2 b mi khc nhau: b th nht
xc nh s ng nht ca PCR v b th hai nhm vo cc trnh t ADN d tha. PCR a thnh
phn cng c th c s dng trong phng th nghim vi sinh lm sng vi cng on d cho mt mu
xt nghim cho vi sinh vt khc nhau trong mt ng PCR.
Khi tin hnh PCR a thnh phn cn phi xem xt mt s yu t trong thit k on mi. Cc mi
phi c nhit lai tng t nhau. S khc bit 10oC trong nhit lai gia cc b mi c th lm cho
cc sn phm khuch i rt khc nhau hay to s khuch i khng th pht hin c ca sn phm
ny hay sn phm khc. Cc on mi c s dng cng c thit k sao cho cc sn phm khc
nhau v kch thc mi sn phm c th c xc nh nhng khng qu khc nhau cho s
khuch i ca mt ADN ch c u tin hn so vi s khuch i ca ADN ch khc. PCR a thnh
phn c cc ADN ch khc nhau nhiu v kch thc thng u tin khuch i cc ADN ch ngn
trc cc ADN ch di, kt qu l c s khc nhau v s lng cc sn phm c khuch i. Tuy
nhin, s khc nhau ng k v nhit lai hay chiu di ADN ch l khng th trnh khi. Ngi ta
c th iu chnh quy trnh thc hin lm cn bng phn ng v t c s lng sn phm tng
ng nhau hoc iu chnh nng mi cn bng vi mt phn ng a thnh phn.
9.5.7.4. PCR ca ARN
ARN ca retrovirus c ly trch v c chuyn i thnh cADN bi enzym sao m ngc enzym
phin m ngc, cADN mi c dng lm khun mu cho PCR. Tuy nhin, cc phn ng nh enzym
phin m ngc kh thc hin v vic s dng cc enzym khng chu c nhit trn 42oC l mt bt
li v xy ra s lai khng nghim ngt. V vy, s phin m ngc km hiu qu l mt tr ngi lm cho
s pht hin cc ARN ch km nhy v km c hiu.
Ngi ta tm ra ADN polymerase chu nhit c hot tnh enzym phin m ngc l ADN
polymerase ti t hp c trch t vi khun chu nhit Thermus thermophilus (Tth pol). ADN
polymerase chu nhit ny hot ng hu hiu khi c s hin din ca ion Mn2+. Hn hp phn ng ca
PCR ca ARN bao gm: Tth pol, 2 loi mi oligonucleotide (mt c s dng cho s tng hp
cADN, v c 2 c s dng cho PCR), ion mangan dng MnCl2, v tt c cc thnh phn khc cn
cho s phin m ngc v PCR. My chu k nhit c un nng trc n 70oC, v nhng thnh phn
phin m ngc c trong 15 pht. Sau phn ng nh enzym phin m ngc, hn hp phn ng
c nng nhit ln 95oC lm bin tnh phc hp ARN - ADN. PCR sau c bt u vi 2
chu k 95oC trong 15 pht v 60oC trong 20 giy, tip theo l 38 chu k 90oC trong 15 giy v 60oC
khong 20 giy. S phin m ngc c thc hin 70oC do vy c hiu v nhy ca vic pht
hin ARN ch cao.
9.5.8. Cc lnh vc ng dng ca PCR
Trong nghin cu khoa hc, PCR gip cho vic xc nh trnh t nucleotid ca cc on ADN c
nhn ln; c th s dng PCR tch dng nhng on ADN c hiu, mc d trong nhiu trng hp
tch dng khng cn n PCR; n gip pht hin t bin, nghin cu mARN hoc to cc t bin gen,
cho php phn tch lin kt gen t nhng t bo ring l; gip nghin cu qu trnh tin ho mc phn
t. Thm ch gip phc hi nhng gen trong cc c vt tn ti cch y hng chc triu nm.
Trong chn ging vt nui v cy trng, c bit i vi ng vt v thc vt bc cao, vn trc y

vic la chn cp cha m lm ging l rt kh khn, cn nhiu nm, th nay PCR cho php thc hin ch
trong vi ngy, thm ch vi gi vi hiu qu rt cao.
Trong y hc, PCR c th chn on chnh xc cc bnh nhim trng t virut n vi khun v cc
bnh do nm, k c HIV - AIDS, chn on sm v theo di iu tr ung th...
Trong t vn di truyn y hc, PCR cho php chn on nhanh, chnh xc cc bnh di truyn k c
chn on trc sinh gii tnh v cc d tt bm sinh c th c t khi thai mi c 8 tun tui v iu
quan trng l khng cn chc i, ch cn ly mt git mu ngoi vi ca m lm mu th nghim.
Trong khoa hc hnh s, PCR l khng th thiu, n gip chn on nhanh, chnh xc th phm ch
t mt vt mu kh, mt si tc hoc mt sinh phm no m th phm li trn hin trng. Ngoi
ra k thut ny cn cho php xc nh chnh xc quan h huyt thng cha - con, ng - chu v.v... cng
ch trong vi gi.
Trong bo v mi trng, vic xc nh mc nhim sinh hc c th thc hin rt hiu qu v

nhanh chng bng phn ng PCR.
Nhng ng dng thc tin a dng ca phn ng chui tng hp ADN tht v cng tn. Nu nh
nm 1985 mi c 3 cng trnh c cng b v PCR th 5 nm sau k thut ny c s dng trong
hng nghn phng th nghim trn khp th gii. Ngay nc ta, PCR cng v ang c dng trong
nhiu trng i hc, vin nghin cu v ngy cng tr nn ph bin. Phn ng chui tng hp ADN
tht s a li mt cuc cch mng trong lnh vc ng dng thc t ca sinh hc phn t. Ngy nay
c th ni mi lnh vc nghin cu sinh hc u s dng PCR.
Tm tt
Chit tch v tinh ch ADN ng vai tr quan trng trong cc nghin cu sinh hc phn t. Cc
bc chnh chit tch ADN l ph v t bo, loi b protein, v ta acid nucleic. Tinh ch ADN c
th p dng k thut siu ly tm hoc sc k. Nng ADN c th o bng quang k bc sng 260
nm, kch thc tng i ca ADN c th xc nh bng k thut in di trn gel i chiu vi thang
kch thc bit.
Cc thao tc ct, ni v lai ADN l cc thao tc c bn ca k thut di truyn phn t. Trnh t ADN
c th xc nh bng phng php ho hc (Maxam v Gilbert) hoc phng php enzym (Sanger v
cng s). Phng php Sanger c ci tin n gin hn v c th t ng ho.
Phng php PCR s dng enzym ADN polymerase v my lun nhit cho php khuch i in vitro
acid nucleic ln gp t ln sau 30 - 40 chu k ch trong vi gi. K thut ny c nhiu u im nh n
gin, nhanh v nhy nn c ng dng rng ri trong nghin cu khoa hc, chn on, khoa hc hnh
s, chn ging...
CU HI
1. Cch no khng dng tinh ch and?
a) Sc k i lc.
b) Sc k lc gel.
c) Sc k trao i ion trn vi ct.
d) Sc k lng hiu nng cao.
e) Sc k kh
2. Tc in di khng ph thuc:
a) Kch thc phn t ADN
b) Cu dng ca ADN
c) Nng ADN
d) Nng gel
e) in th s dng
3. Enzym ct gii hn loi c ng dng nhiu trong k thut ti t hp di truyn
a) I b) II
c) III d) I v III
e) II v III
4. Yu t nh hng n s lai ho:
a) Nng ADN b) Nhit v thi gian phn ng
c) di ca cc trnh t d) Lc ion
e) Tt c
5. c im khng thuc phng php nh trnh t ca Sanger:
a) X l ho hc chuyn bit lm bin i c trng mt loi nucleotid
b) S dng ADN polymerase
c) Nucleotid c nh du.
d) Phn ng tin hnh trong bn phn on
e) C s dng dideoxynucleotid
6. Cht lm gim nhit bin tnh ca ADN trong PCR
a) Formamid b) MgCl2
c) DMSO d) EDTA
e) a v c
7. Mi trong phn ng PCR
a) on ADN ngn, mch n
b) C trnh t b sung vi ADN khun ti im u sao chp
c) Di t 6 - 30 nucleotid
d) L oligonucleotid
e) Tt c
8. Tnh nhit "chy" ca on mi nhm xc nh nhit thch hp
a) Bin tnh mi
b) Mi gn vo khun
c) Tng hp t khun trn gen
d) Mi khng gn b sung vo nhau
e) Mi gn vo cc on khc nhau
9. Chn chu k nhit PCR da vo 2 yu t l kch thc ca khun v
a) tinh khit ca khun b) Nng ca khun
c) Kch thc mi d) Trnh t mi
e) c v d
10. PCR t:
a) Da trn nguyn tc ca phn ng PCR
b) L hai PCR lin tip, s dng hai cp mi "ngoi" v "ni"
c) C nhy v chuyn bit cao hn PCR thng
d) ng dng trong chn on
e) Tt c ng
P N
Bi 1. Nhp mn sinh hc phn t
1. b 2. e 3. c 4. e 5. a
Bi 2. Sao chp ADN
1. d 2. b 3. d 4. e 5. e
6. e 7. a 8. b 9. b 10. b
Bi 3. Cc loi ARN
1. e 2. a 3. a 4. d 5. b
6. b 7. e 8. c 9. d 10. e
Bi 4. S phin m v m di truyn
1. e 2. e 3. e 4. c 5. e
6. b 7. c 8. c 9. e 10. a
Bi 5. Sinh tng hp protein
1. e 2. d 3. a 4. a 5. b
6. d 7. e 8. e 9. d 10. c
Bi 6. iu ho hot ng gen
1. e 2. e 3. d 4. b 5. a
6. e 7. d 8. e 9. b 10. b
Bi 7. B gen t bo nhn tht
1. b 2. d 3. e 4. e 5. e
6. e 7. e 8. e 9. e 10. e
Bi 8. t bin gen
1. c 2. e 3. e 4. d 5. d
6. e 7. e 8. d 9. c 10. a
Bi 9. Cc phng php phn tch ADN
1. e 2. c 3. b 4. e 5. a
6. e 7. e 8. b 9. e 10. e
TI LIU THAM KHO

1. Benjamin Lewin, Genes VIII. Prentice - Hall, 2004.
2. H Hunh Thu Dng, Sinh hc phn t. NXB Gio Dc, 1997.
3. James Swarbrick, Pharmaceutical Gene Delivery Systems. Marcel Dekker Inc. New York, 2003.
4. L nh Lng. Nguyn l k thut di truyn. NXB Khoa hc K thut, 2001.
5. Phm Thnh H. Di truyn hc. NXB Gio Dc, 1998.
6. Richard A. Harvey, Pamela C. Champe. Biochemistry. 3rd ed. Lippincott Williams and Wilkins,
Baltimore, 2005.
7. Richter, J. D. in Translational Control of Gene Expression (Hershey, J. W. B. , Mathews, M. B. ,
and Sonenberg, N., eds), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2000.
8. Sambrook, J., Fritsch, E., v Maniatis, T. Molecular Cloning: A Laboratory Manual, 2 Ed., Vol. 3,
Cold Spring Harbor Press, Cold Spring Harbor, New York, 1989.
9. Smith C. A. and Wood E. J. Molecular biology and Biotechnology. Chapman & Hall, 1991.
10. Tom Strachan and Andrew P. Read. Human Molecular Genetics. Bios Scientific Publishers Ltd,
1996.
Chu trch nhim xut bn:

Ch tch HQT kim Tng Gim c NG TRN I
Ph Tng Gim c kim Tng bin tp NGUYN QU THAO
Chu trch nhim ni dung:

Ch tch HQT kim Gim c Cng ty CP Sch H DN
TRN NHT TN
Bin tp v sa bn in:
TRN NGC OANH
Trnh by ba:
BI QUANG TUN
Ch bn:
THI SN
SINH HC PHN T
M s: 7K721M7 DAI
In bn, (Q: ) kh 19 27 cm, ti
S KKH xut bn: 770 2007/CXB/3 1676/GD
In xong v np lu chiu thng 12 nm 2007.

Sinh Hoc Phan Tu

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New Page 2 Page 1 of 203

Nhng ngi bin son:

Tham gia t chc bn tho:

Bn quyn thuc B Y t (V Khoa hc v o to)

770 - 2007/CXB/3 - 1676/GD M s: 7K721M7 - DAI

Sch SINH HC PHN T c Hi ng chuyn mn thm nh sch v ti liu dy - hc

B Y t xin chn thnh cm n cc tc gi v Hi ng chuyn mn thm nh gip hon thnh

Ln u xut bn, chng ti mong nhn c kin ng gp ca ng nghip, cc bn sinh vin

Sch xut bn ln u nn kh trnh khi thiu st. Cc tc gi rt mong nhn c s gp ca

NER Nucleotide excision repair

ssADN Single stranded DNA

BNG I CHIU THUT NG VIT - ANH

ADN b sung Complementary DNA

ADN si i Double - stranded DNA

ADN si n Single - stranded DNA

ADN vi v tinh Microsatellite DNA

Alkapton niu Alkaptonuria

ARN nhn khng ng nht, hnARN Heterogenous nuclear RNA

ARN nhn nh, snARN Small nuclear RNA

ARN polymerase ph thuc ADN DNA - dependent RNA polymerase

ARN ribosom Ribosomal RNA

ARN t bo cht nh, scARN Small cytoplasmic RNA

ARN thng tin Messenger RNA

ARN vn chuyn Transfer RNA

Base ng ng Base analogue

Bnh hng ban lupus Systemic lupus erythematosus

Bnh lon dng c Duchenne Duchenne muscular dystrophy

B ba kt thc Stop codon

B suy gim Attenuator

Bong bng sao chp Replication bubble

Chc ba sao chp Replicating fork

Cht d nhim sc Heterochromatin

Cht d nhim sc c cu Constitutive heterochromatin

Cht hot ho Activator

Cht nguyn nhim sc Euchromatin

Cht nhim sc Chromatin

Cht siu c ch Superrepressor

Cht c ch Inhibitor, repressor

Chip ADN DNA microarray

Chui xon kp Double helix

Chuyn v nghch Retrotransposition

C ch ln vng Rolling circle mechanism

Cng ngh di truyn Genetic engineering

Cng ngh sinh hc Biotechnology

Cng ngh nano sinh hc Bionanotechnology

Du n ADN DNA fingerprint

D n b gen ngi Human genome project

a kim sot Multiple control

u dnh Cohesive end

im gc sao chp Origin of replication

in di gel Gel electrophoresis

Pulse field gel electrophoresis

iu ho dng Positive control

nh kiu gen Genotyping

on chn Insertion sequence

on Okazaki Okazaki fragment

n v sao chp Replicon

t bin cm ng Induced mutation

t bin cng ngha Sense mutation

t bin lng Silent mutation

t bin lch ngha Missense mutation

t bin t nhin Spontaneous mutation

t bin v ngha Nonsense mutation

Enzym ct gii hn Restriction enzyme