CHNG IV

KIM SOT PHIN M V HU PHIN

M QU TRNH BIU HIN GEN M QU TRNH BIU HIN GEN
NI DUNG
Biu hin gen v kim sot biu hin gen
Kim sot biu hin gen sinh vt nhn
s s
Kim sot biu hin gen sinh vt nhn
chun
KIM SOT BIU HIN GEN
S kim sot biu hin gen l s kim sot mc ,
thi im biu hin ca gen nhm m bo cho s tn
ti, pht trin hi ha ca t bo, c th p ng li s
thay i ca mi trng bn trong v bn ngoi c th
S kim sot biu hin gen l s kim sot mc ,
thi im biu hin ca gen theo mt chng trnh pht
trin chuyn bit, hng n vic chuyn bit ha tng
loi t bo vo cu trc, chc nng ring trong qu trnh
pht trin ca c th.
IU HA BIU HIN GEN
iu ha biu hin gen trong qu trnh pht trin phi
rui dm
S biu hin gen c kim sot cht ch. Nhng gen
khc nhau c hot ha biu hin nhng t bo khc
nhau.
S biu hin gen c kim sot cht ch. Nhng gen
khc nhau c hot ha biu hin nhng t bo khc
nhau.
Histochemical localization of GUS expression under control of the Atndt1 and
Atndt2 promoter in A. thaliana.
iu ha biu hin gen nm men cc iu kin mi
trng v kiu gen khc nhau
C CH KIM SOT BIU HIN CA GEN?
Thuyt trung tm ca sinh hc phn t
Cc cp
iu
ha kim
sot biu sot biu
hin gen
Kim sot biu hin gen sinh vt nhn s
c im cu trc gen, qu trnh phin m sinh vt
nhn s
Kim sot biu hin gen mc phin m
-iu ha m tnh: ngn cn phin m
-iu ha dng tnh: tng cng phin m -iu ha dng tnh: tng cng phin m
S suy gim biu hin gen: iu ha bng phin m
d- tng hp tryptophan
C ch iu ha chung: p ng ca E.coli vi sc
nhit- s iu ha biu hin ca tp hp nhiu gen khc
nhau.
ARN polymerase l enzyme ch yu cho phin m
ARN polymerase tham gia vo ba giai on phin m:
-M !u: tiu n v" # + enzyme li ($$%%) bm vo
promoter, m xo&n ADN, b&t !u phn ng polymer ha
cc base b sung vi ADN khun
-Ko di: d"ch chuyn kh'i promoter, gii phng tiu
n v" #, phn ng polymer ha n v" #, phn ng polymer ha
-Kt thc: ARN polymerase chuyn ti tn hiu kt thc
Ph( thuc Rho: nhn t Rho nhn ra trnh t trn
mARN, bm vo n v tch n kh'i ARN
polymerase
Khng ph( thuc Rho: cu trc k)p tc c hnh
thnh gy dng phin m
Cc trnh t iu ha
Cc promoter
- i lc ca ARN polymerase vi promoter ca gen
- v" tr nhn bit v bm ca ARN polymerase
TTGACA (vng 35) v TATAT (vng 10- hp
Pribnow) Pribnow)
- promoter mnh c hiu sut hot ng cao vi
mc phin m 2s/l!n, ngc li 10 pht/l!n
iu ha thng qua nhn t #
iu ha thng qua nhn t kt thc, nhn t phin m
d, nhn t khng kt thc
Operon
Cc gen c t chc thnh cc n v" phin m gi
l operon, vi mt promoter duy nht, hnh thnh nn
sn phm l mt ARN polycistronic.
Kim sot biu hin gen sinh vt nhn s
M hnh Lac Operon
1961: Monod v Franois Jacob pht hin ra m hnh operon khi nghin
cu cc dng t bo E. coli t bin gen s dng lactose.
Mt s khi nim
C nh (constitutive)
Thch ng (Adaptive) Thch ng (Adaptive)
Mt s khi nim
C "nh (constitutive)
-Lun biu hin vo mi thi im vi lng c "nh
-Khng c s p ng vi mi trng
Thch ng (Adaptive)
-p ng vi s thay i ca mi trng
c th c cm ng (inducible)
c th b" km hm (repressible)
U
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s

-
g
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No lactose
+ lactose
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b
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y
m
e
s+ argenine
Time
No lactose
L
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o
f

A
r
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b
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s
y
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Time
Cmng (Inducible to sn
phm!p "ng v#i ch$t cm
"ng% th&'ng l( c) ch$t c*a
enzyme
!"mh#m($epressible b+ ,-m
h.mb/i ch$t 0ng ,-mh.m%
th&'ng l( sn phmc12i c3ng
c*a ch14i phn "ng sinh t5ng
h6p
lac
%
&
'
7nco8es -galactosi8ase
7nco8es permease
'
7nco8es transacetylase
9:
http;<<en=>i,ipe8ia=org<>i,i<Lac?operon
9@
Kim sot tiu cc- vai tr ca yu t km hm
iu ha km hm:
- Phin m trn operon lac b" c ch bi yu t km hm l protein
LacI. Khi khng c mt lactose, protein LacI bm vo operator
lm ngn cn phin m
iu ha cm ng:
-Khi c mt cht cm
ng (allolactose, IPTG),
cht ny bm vo yu t
km hm (protein LacI)
lm thay i cu hnh lm thay i cu hnh
cht km hm nn khng
bm vao operator c.
- ARN polymerase bm
vo promoter v b&t !u
qu trnh phin m
Bng cch thc no m Monod-Jacob tm ra
c c ch kim sot ca operon lac?
Cc dng t bin
t bin lacI
-
- Cht km hm khng th bm vo operator , do
khng th gy km hm phin m.
Cc dng t bin
t bin lacS
-
- Cht cm ng khng th bm vo cht c ch, do
cht c ch lun lun gy km hm phin m
t bin O
c
- Cht km hm khng th nhn din v bm vo vng
operator (do trnh t vng operator b" thay i), qu
trnh phin m di*n ra lin t(c
Bng chng thc nghim xc nh protein km hm: Th
nghim PaJaMo (Pardee, Jacob v Monod)
a lacI
+
lacZ
+
vo t bo t
bin lacI
-
lacZ
-
%-galactosidase
c tng hp
trong vng 1-2 h trong vng 1-2 h
!u ri dng li
Nu b sung
cht cm ng %-
galactosidase li
tip t(c c tng
hp
Bng chng thc nghim chng minh cht c ch bm vo
vng operator
Th nghim xc nh vng protein iu ha bm vo ADN-DNA
footprint
iu ha kiu trans, kiu cis
Yu t iu ha kiu trans: c th tc ng biu hin
gen nm bt k v" tr no trong h gen. Loi ny
thng l protein.
Yu t iu ha kiu cis: ch+ gy nh hng n biu
hin ca gen nm g!n k n trn cng si ADN. Loi
ny thng l on trnh t ADN.
iu ha kiu trans, thng l protein
iu ha kiu cis, thng l ADN
Laci
+
: to cht km hm c kh nng bm vo vng
operator hoc cht cm ng
Laci

: to cht km hm khng c kh nng bm vo

vng operator
AB
Laci
S
: to cht km hm c kh nng bm vo operator,
nhng khng bm c vo cht cm
Tnh tri ca alen: i
S
> i
+
> i

Hy d on kt qu th nghim
iu ha kiu trans, kiu cis
Protein LacI iu ha kiu trans.
Operator iu ha kiu cis.
Kim sot tch cc- vai tr ca yu t hot ha
iu ha tch cc lm
tng biu hin ca cc
gen lacZ, lacY v lacA.
cAMP (cyclic
adenosine
monophosphate) bm monophosphate) bm
vo CRP (cAMP
receptor protein) khi
nng glucose thp.
CRP bm vo vng
iu ha, tng cng
hot tnh ca ARN
polymerase ti promoter
lac.
Cch thc protein iu ha bm vo ARN polymerase
CRP-cAMP khng ch+ l cht iu ha tch cc ca
operon lac, m cn ca nhiu h thng khc nh operon
gal, operon ara.
,c ch qu trnh d" ha: khi mi trng c mt
glucose v mt s ng khc (lactose, galactose,
arabinose), t bo vi khun ch+ s d(ng glucose v arabinose), t bo vi khun ch+ s d(ng glucose v
km hm hot ng ca cc operon lin quan n chuyn
ha cc loi ng khc. Hin tng ny c gi l c
ch qu trnh d ha( catabolite reppression).
Nh vy, C 2 c ch iu ha operon lac: tiu cc v
tch cc.
Cu h'i: S biu hin gen ca operon lac t cc i Cu h'i: S biu hin gen ca operon lac t cc i
khi no?
Khi mi trng c lactose v thiu glucose. Trong iu
kin ny, cht cm ng bm vo cht c ch lm cht
ny khng c kh nng bm vo operator gy km
hm phin m; trong khi phc hp CRP-cAMP bm
vo v" tr g!n promoter h- tr ARN polymerase b&t
!u phin m. !u phin m.
iu khin kim sot ca operon ara
AraC ng vai
tr va l cht km
hm, va l cht
hot ha ca hot ha ca
operon ara m ha
enzyme thy phn
arabinose
C ch tng tc ca protein iu ha i vi
ARN polymerase
Cht iu ha tiu cc (cht km hm)
- Chn ARN polymerase ti cc v" tr bm vo ADN
khi !u phin m.
Cht iu ha tch cc Cht iu ha tch cc
- Tip xc vi ARN polymerase lm tng cng kh
nng m !u phin m ca enzyme
Kim sot biu hin- Operon tryp
Operon tryptophan gm 5 gen m ha cho cc enzyme
lin quan n vic tng hp tryptophan.
Khi khng c tryp mi trng, hot ng ca
operon tng
Trypophan ng vai tr tc nhn c ch biu hin Trypophan ng vai tr tc nhn c ch biu hin
operon tryp, gi l cht ng c ch (co-repressor)
S km hm operon tryp i h'i tng tc gia cht
ng c ch v cht c ch
Tryp l sn phm cui cng ca operon tryp, gy km
hm hot ng ca operon c ch phn hi (feedback
inhibition)
Operon tryptophan- tryp ng vai tr l cht !ng c ch
Kim sot theo c ch phin m d-(Attenuation)
Cc yu t sigma (") thay th
- Ph!n ln gen prokaryote c phin m bi ARN
polymerase mang yu t
70
. Tuy vy, nhiu gen khc
c phin m bng cc yu t khc. Qua , b my
phin m c huy ng n cc gen vo cc thi im
khc nhau.
- Vic s d(ng cc yu t thay th l mt c ch lp
trnh s biu hin ca gen.
V d(:
32
l yu t hot ng khi c tn hiu sc nhit

54
yu t hot ng cc gen chuyn ha Nit
Cc yu t sigma (") thay th nhit cao