R

O
AM
S,
14
/e
I
E
E

S
T

C
G

N
CYTOGENETIC DISORDERS ƒƒ Performed on nucleated cells e.g.
Blood -WBCs, BM, semen, vaginal epithelial cells tooth G
Exact multiple 3n, 4n, 5n (2n is normal) pulp, hair root, m/s, skin, mucous membrane E
D/to abnormal number of chromosomes

Euploid

N
ƒƒ Uses RFLP or repeat sequence DNA to establish pattern of
E
DNA for individual.
Not in Non- Occurs when a T
exact disjunction homologous pair of I
multiple chromosome fails to CYTOGENETIC TECHNIQUES C
disjoin at 1st meiotic
S
div Karyotyping
(2n-1), (2n +1)
ƒƒ Sample of cells are fixed and stained in metaphase to
Anaphase One homologous
create light and dark bands and the picture of chromosome
Aneuploid

lag chromosome in meiosis

/One chromatid in is analyzed.
mitosis lags behind ƒƒ Typically used to detect germline aneuploidies. Done in
dividing cells only.
Deletion Interstitial (M/c type of deletion)
Terminal Rapid Methods

/e
Ring chromosome
D/to abnormal structure of chromosomes

Sample DNA can be used in interphase or metaphase spread

Inversion Paracentric of chromosome.

14
Pericentric ƒƒ Interphase FISH
Isochromosome 1 arm of chr is lost & other is °° Fluorescence in situ hybridization is performed very
duplicated i.e. 2 short arm or 2 long rapidly (<24 hrs). Can be done in non-dividing cells.
arm
°° Used to detect subtle microdeletions, complex
Translocation Reciprocal Mutual transfer of translocations, telomere alterations and specific
S,
a segment of chr
rearrangements (DNA sequences on chromosomes).
to another non
-homologous chr. °° Used for species identification, genetic counseling.
Balanced, non- °° Most useful apparatus to detect BCR-ABL fusion gene
AM

balanced type is FISH.

Robertso- Translocation b/w 2 ƒƒ Chromosomal painting
nian acrocentric chr.  Entire chromosome can't be seen simultaneously.
e.g Down's
ƒƒ Spectral karyotyping
 Entire chromosome can be seen.
O

High-yield Points
ƒƒ Micro array
44 Incidence of chromosomal anomalies is 0.4% in live births.  A DNA microarray (a/k/a DNA chip or biochip) is a
R

44 All autosomal monosomies are fatal while sex chromosomal collection of microscopic DNA spots (snapshot) attached
monosomies & trisomies are compatible with life.
to a solid surface. Used to measure the expression levels of
44 Trisomies are m/c numeric abnormalities while deletions are
large numbers of genes simultaneously. Rapid & accurate
m/c structural abnormalities.
44 Most trisomies occur d/to meiotic non-disjunction whereas
generation of gene expression.
trisomy 7 occur d/to mitotic non-dysjunction. ƒƒ Comparitive genomic hybridization (cGH array)
44 Common trisomies are → 21 (commonest), 18 and 13. CGH or CMA compares the content of DNA in normal
& tumour cell. Detects only unbalanced chromosomal
CHROMOSOMAL ANALYSIS changes. Balanced translocations or inversions are not
detected.
DNA Fingerprinting ƒƒ Single stranded conformation polymorphism (SSCP)
Can detect point mutation.
ƒƒ Given by Alec Jaffrey in 1984.
[Remember that Fingerprinting/dactylography, which is a ƒƒ Other techniques:
different entity, was given by Francis Galton] MAPH : Multiplex Amplification & Probe Hybridization.
ƒƒ Was a serendipity. MLPA:Multiplex Ligation Dependent Probe Amplifican.
MALDI : Matrix Assisted Laser Desorption & Ionization 213
ƒƒ Analyze short tandem repeat sequences.
 FRAP : Fluorescence Recovery After Photobleaching
G (FRAP) is used for structure of membranes (like Fluid High-yield Points
E mosaic model in the past).Movement of protein from 44 Prenatal d/g of neural tube defects is not possible by Chorionic
N nucleus to cytoplasm is seen. villous sampling.
E ƒƒ Linkage analysis of the genes is done by → Chromosomal 44 Muscle dystrophy cannot be diagnosed by CVS or amniocentesis.
T walking. Muscle biopsy is diagnostic.
I 44 Prenatal d/g of hemophillia is based on linkage analysis. Genetic
C N Important negative points: Cytogenetic techniques linkage analysis c/b performed by RFLP
S ŽŽ FISH is NOT used to introduce genome into bacteria or 44 Microsatellite sequence is short sequence (2-5) repeat DNA.
into target cell.
ŽŽ MALDI is NOT used to diagnose subtelomeric
rearrangements of genes. MENDELIAN INHERITANCE
ŽŽ Dideoxyribonucleotides are NOT required in PCR.
XR
ƒƒ Manifests only in males. E.g. hemophilia which manifests
4.6 GENETIC BASIS OF DISEASES only in boys

/e
ƒƒ All females are carrier (do not manifest the disease)
ƒƒ Son of female carrier, 50% are affected
PRENATAL DIAGNOSIS
Daughter of female carrier, 50% are carrier

14
ƒƒ Around 50% of abortions, 50% of congenital anomalies,
50% of mental retardation, 50% of deafness, 50% of XD
blindness and 15% of cancers have genetic basis. ƒƒ No male to male transmission
ƒƒ Sample of cells obtained by amniocentesis or CVS is used ƒƒ Only females affected
for prenatal diagnosis. Pregnant women of 35 or + are
S,
candidate for prenatal diagnosis as risk of aneuploidy is AD
higher in woman of age 35 or more. ƒƒ Vertical inheritance
ƒƒ Enzyme replacement possible for Gaucher's d/s.
AM

ƒƒ 50% chance of inheriting a d/s

ƒƒ Method of choice for prenatal diagnosis of hemophilia →
direct mutational analysis based on PCR.
ƒƒ Down's syndrome is a/w trisomy 21. In 95% the extra
chromosome is of maternal origin, in 4% cases this extra
chromosome is derived from Robertsonian translocation
O
ƒƒ Variable expressivity/ incomplete penetrance is seen in AD
disorders.
ƒƒ AD disorders are manifested in the heterozygous state, so
at least one parent of an index case is usually affected.
ƒƒ When an affected person marries an unaffected person --

of 21q to another acrocentric chromosome t (13,21). In 1% Each child has 50% chance of having d/s.
cases patients are mosaic. ƒƒ If both parents have d/s (heterozygous) --- Chances of
R

Prenatal Diagnosis for Genetic Disorders having an unaffected baby are 25%
ƒƒ If both parents are homozygous --- all the children will be
Several single gene disorders are diagnosed prenatally by
affected [ 0% children will be unaffected]
DNA analysis
ƒƒ AD disorders: AR
°° Huntington's ds
ƒƒ Horizontal inheritance. Transmitted in many siblings.
°° Myotonic dystrophy ƒƒ 25% chance of affecting a child
°° Neurofibromatosis ƒƒ 50% are carrier
ƒƒ AR disorders:
ƒƒ Complete penetrance is seen.
°° Sickle cell anemia ƒƒ The d/s is expressed only in homozygotes.
°° Cystic fibrosis ƒƒ The parents of an affected child are asymptomatic.
°° Thalassemia α & β ƒƒ Each child born to heterzygous parent has 25% risk of
°° Tay-sachs d/s being affected.
°° PKU (Phenylketonuria) Children of an affected parent will themself not be affected,
ƒƒ X-linked disorders: unless the other parent is a heterozygous or is obviously also
°° Hemophilia A&B affected.
214
°° Duchenne type m/s dystrophy
Approach Inheritance Pattern through Pedigree ƒƒ If there is no sex bias → look for affected parents
G
Condition AD AR XR Mit E

% of 50% if 1 0 % if 1 0 % of 0 % if N
offspring heterozygous carrier parent sons if dad is Parents affected Parents not affected
E
affected parent & 1 & 1 parent father is affected
unaffected homozygous affected
T
All generations have In 2nd generation 50% I
parent; unaffected;
50% of 100% if
affected individuals & in 3rd generan 25% C
75% if both 25% if both sons if mom is (parents and all generations individuals affected S
heterozygous heterozygous mom is a affected are automatically dominant)
affected carrier parents carrier;

parents
100%of
The disorder is AD The disorder is AR
100% if 1 50% if 1 sons if
homozygous homozygous mom is
affected affected parent affected
parent &1 Some Examples of Inheritance
heterozygous

/e
carrier
parent

14
Inheritance Vertical Horizontal Criss cross
pattern
Male & Yes Yes No (Males Yes
female affected
S,
offsprings oftenly)
affected at
the
same rate Fig.: AD inheritance
AM

Allele Either Either Sons = Mom

inherited Mom only only
from which Daughters
parent(s)? = either
Can be No Yes Yes No
inherited
O

from
unaffected
parents?
R

To Summarize
ƒƒ First of all in a generation look for..if only one sex is
affected... Fig.: AR inheritance
Condition a: only males affected
(Mama - Bhanja effect) only males with maternal uncles
affected → The disorder is X LINKED RECESSIVE.
E.g. hemophilia which manifests only in boys.
ƒƒ Condition b: only females affected
(Baap - beti ka pyar) affected males transmit only
to their daughters → The disorder is X LINKED
DOMINANT.
(Becoz males do not survive).

215
Fig.: X-linked recessive inheritance
 X-Linked Recessive X-Linked Dominant
G
•• Agammaglobulinemia •• Alport syndrome
E
(Bruton's) •• blood group Xg
N •• Adult PCKD – colour •• Fabry's Ds
E blindness •• Familial hypophosphatemia
T •• Color blindness (VDRR, Vitamin D resistant
I •• CGD (Chronic rickets)
granulomatous d/s) •• Type 3 VDDR, HPDR
C
•• Duchenne's and Becker's •• G-6-P D def. (incompletely
S MD dominant expression)
Fig.: Mitochondrial inheritance •• DI •• Incontinentia Pigmenti
•• G-6-P D def. (incompletely
dominant expression)
High-yield Points •• Hemophilia A &B
•• Ocular albinism
44 In some AD disorders(e.g. Osteogenesis imperfecta) •• Ornithine transcarbamylase
phenotypically normal parents have >1 affected child. This is deficiency
exception to normal mendelian inheritance. This is d/to germline •• Retinitis pigmentosa
•• Fragile X syndrome

/e
or gonadal mosaicism.
44 In AD disorders some individual inherit the mutant gene but •• Fabry's, Hunter's d/s
phenotypically normal. This is d/to incomplete penetrance. 50%
penetrance means 50% individual who carry the gene express

14
Polygenic (multifocal) Inheritance is seen in
the trait.
44 If trait is seen in all individuals carrying the mutant gene but ƒƒ DM
expressed differently among individuals, it is called variable ƒƒ Essential hypertension
expressivity. Seen in neurofibromatosis type I. ƒƒ Cleft lip/palate
ƒƒ Autism
S,
MENDELIAN DISORDERS [Single gene ƒƒ Cancers
disorders]
AD AR High-yield Points
AM

•• Achondroplasia Most IEMs (inborn errors of 44 Autosomal Codominance (codominant Inheritance) is seen in →
•• Acute intermittent porphyria metabolism) HLA, Blood group Ag.
•• Adult PCKD •• Albinism 44 Remember most of the inborn errors of metabolism are AR
•• BRCA1/2 breast cancer •• Alkaptonuria disorders except → Hunter's, Fabry's ds, G6PD def. (XR),
•• MODY ( Maturity onset •• Agammaglobulinemia (swiss
Bruton's agammaglobulinemia, ocular albinism (XR), familial
O

diabetes of youngs) type)

•• Familial - •• Ataxia telangiectasia hypercholesterolemia (AD).
ƒƒ Hypercholesterolemia •• a1Anti-trypsin deficiency 44 Disorders with prefix 'hereditary' or 'Familial' are usually
ƒƒ Hypertrophic CMP •• Βeta thalassemia AD disorder e.g. FPC (familial polyposis coli, Familial
R

ƒƒ Dilated cardiomyopathy •• Cystic fibrosis hypercholesterolemia, Hereditary spherocytosis).

ƒƒ Colonic polyposis
•• Hereditary -
ƒƒ HNPCC
ƒƒ Hemorrhagic
telangiectasia
ƒƒ Spherocytosis (HS)
•• Huntington's Chorea (HD)
•• Hyperlipoproteinemia
1,2,3,4
•• Von Willebrand disease
•• Marfan syndrome
•• Mytonic dystrophy
•• M/s dystrophy, limb girdle -1 •• PKU
•• Neurofibromatosis
•• Otospongiosis /otosclerosis •• Hemochromatosis
•• Osteogenesis imperfecta
•• Peutz Jeghers syndrome
216
•• Polydactyly
•• Retinoblastoma
•• vWD
•• Congenital erythropoietic
porphyria (CEP) Epigenetic Effect
•• CAH, 21-Hydoxylase def.
•• Friedreich's ataxia ƒƒ Meiotically and mitotically heritable changes in gene
•• Fanconi's syndrome expression not a/w DNA sequence alterations are referred
•• Glycogenosis, Gaucher’s ds., to as epigenetic effects.
PK deficiency ƒƒ Role in cancer, mental retardation & hematologic disorders
•• Hirschsprung's disease
and ?aging.
•• Lysosomal storage ds
•• M/s dystrophy , limb girdle
Polymorphism Mutation
-2
•• Maple Syrup Urine ds •• Definition Clinically harmless Infrequent,
variation of DNA that potentially harmful
•• Sickle cell anemia does not affect the
phenotype
•• Wilson's ds
•• Affects Single phenotype, Single mutation →
•• Homocystinuria
single locus multiple end effects
•• Wilson's d/s
(Pleiotropism)
ƒƒ Gene polymorphism °° Chronic progressive external ophthalmoplegia
Gene expressing variability (E.g. skin colors). Clinically Pearson's syndrome G
°°
harmless variation of DNA that does not affect the Inherited myopathies(MMC, ADMIMY syndrome) E
°°
phenotype. MERRF N
°°
ƒƒ Genetic pleiotropy (Pleiotropism) °° NARP and Leigh d/s E
Defect in a single gene causes multiple unrelated problems. T
E.g. Mosaicism I
°° In Alport syndrome mutation of a5 collagen chain ƒƒ Presence of >2 distinct cell lines in the tissue of an C
affecting kidney(hematuria),eye (lenticonus) & ear individual. (E.g +nce of 47 XXX & 45,X in the same S
(SNHL). person).
°° Phenyl ketonuria
During mitotic division
ƒƒ Genetic heterogenecity (Heterotropism)
Multiple mutations causing → single end effects or trait

(E.g. Childhood deafness, Hereditary spherocytosis
1 cell gets 1 extra chromosome Other cell gets 1 less chr
(mutations in spectrin, ankyrin, spectrin all leading to HS).
ƒƒ Antegrade pleiotropy
47 XXX 45,X

/e
Single mutation causing → two opposite effects (E.g. In
p53 gain mutation → No tumour formation (beneficial (Turner)
effect) & growth retardation (harmful effect). ƒƒ Chromosomal mosaicism from non-disjunction results in

14
Turner's syndrome.
ƒƒ In somatic mosaicism d/to mutations (non-disjunction
EXCEPTION TO SIMPLE MENDELIAN
occurs in mitosis) E.g. in Gsa results in the McCune-
INHERITANCE
Albright syndrome.
ƒƒ Germline/ gonadal mosaicism >1 cell line derived from a
S,
Mitochondrial inheritance
single zygote. Phenotypically normal , can transmit the d/s
ƒƒ All mitochondria are contributed by the ovum during
to offsprings through mutant gonads. E.g. Osteogenesis
zygotic formation – it is transmitted by maternal non-
imperfecta, tuberus sclerosis.
AM

mendelian inheritance”.
ƒƒ Inherited mitochondrial disorders are transmitted in a Genomic Imprinting (GI)
matri-linear fashion. All children from an affected mother
ƒƒ Selective inactivation of a genes on either paternal
(Mutations in mtDNA) will inherit the d/s. Since only
daughters pass on the mitochondria, mothers who only chromosome or on maternal chromosome (autosome). It
O

have sons end the transmission of that mitochondrial type. leads to preferential expression of functional allele of one
of the parent. 70- 80% of Prader Willi and Angelman's
Mitochondrial DNA (mt DNA) are d/to genomic imprinting and only 20% are d/to
R

ƒƒ Human mitochondrial DNA (mt DNA) is a circular double
stranded DNA molecule containing 16,569 base pairs.
ƒƒ mt DNA accounts for 1% of cellular DNA.
ƒƒ Encodes for 2 rRNA (16s & 12s), 22 mt tRNA, 13 protein
subunits of ETC.
ƒƒ Transmitted by non-mendelian inheritance.
ƒƒ It contains no (or very few) introns or untranslated
sequence.
ƒƒ Has high mutation rate.
ƒƒ Do not recombine like nuclear chromosomes.
ƒƒ D/s transmitted by mt DNA (mitochondrial diseases)
°° Leber’s hereditary optic atrophy
°° Kearns Sayre syndrome
°° MELAS (Mitochondrial encephalopathy with lactic
acidosis & stroke like features)
unilateral parental disomy (UPD).
[Mnemonic to remember : PaPa -G and Uni-MaP means
in Prader Willi Genomic imprinting (deletion) of Paternal
chromosome is common, while UPD of Maternal
chromosome is seen in Prader Willi]
ƒƒ Other examples of genomic imprinting are ---H.mole,
ovarian teratoma, Russell Silver syndrome, Beckwith
Wiede­mann syndrome (d/to expression of imprinted gene
IGF-II)
Uniparental Disomy (UPD)
ƒƒ When both the copies of chromosomes comes from single
parent and this is k/as “Uniparental disomy”
ƒƒ Maternal UPD occurs in Prader Willi syndrome.
Paternal UPD occurs in Angelman's syndrome. 217
 Features Prader Willi Angelman's ƒƒ Methods of transfer of genetic material in
G bacteria: Transfer methods are classified on the basis of
E Chromosome micro deletion at micro deletion at
agencies which transmit genetic material in bacteria:-
N involved 15q 11-13 15q 11-13
E Mostly d/to Paternal genomic Maternal genomic Methods Definition Agency which
T imprinting (70%) imprinting (80%) of transmits
I Less common UPD (25%) of UPD (2-3%) transfer genetic
C cause 2 maternal of 2 paternal informan
S chromosomes chromosomes Trans- Transfer of a genetic Bacteriophage
Cl/f ↓ Fetal activity (Happy puppets) duction material i.e. a portion (virus)
neonatal hypotonia, Normal at birth, of chromosomal DNA
obesity, mental retardation, (episome or plasmid) from 1
mental retardation, seizures, ataxia, bacterium to another using
small hand/feet, hypotonia bacteriophage
Hypogonadism Trans- Transfer of a portion of Free DNA
formation chromosomal DNA from 1
Atavism bacterium to another using

/e
ƒƒ When a child resembles its grandparents it is k/as atavism. Free DNA.
ƒƒ Best example of atavism is caudal appendages as coccygeal Studied in pneumococci
(Griffith) & H. influenzae
projection.

14
Lysogeny Phage DNA becomes Prophage DNA
X-inactivation (Barr bodies) integrated with bacterial
chromosome. Role in
ƒƒ Inactive X-chromosome is seen as Barr body /sex
conferring toxigenicity in
chromatin in buccal mucosa & epidermis and as Drum diphtheria
stick in neutrophils. Barr body is found in interphase of
S,
Conjugan Donor/male bacterium Sex pili
cell cycle
contacts with another
ƒƒ Number of bar bodies = no. of X – 1 (e.g. in Turner's it is recipient.
0, in Klinefelter's it is 1 & in super female XXX it is 2) First demonstrated in E. coli
AM

K12 strain by Lederberg and

Chromosomal Breakage Syndromes Tatum
Most are inherited as recessively or may be induced by
sunlight, LSD abuse in pregnancy. Examples are ƒƒ Transduction is used as a method of genetic engineering in
ƒƒ Fanconi’s anemia the t/t of some IEMs.
O

ƒƒ Bloom’s syndrome ƒƒ Transfection is infection of a bacterium by the naked

ƒƒ Incognita pigmenti bacteriophage nucleic acid.
ƒƒ Ataxia telangiectasia (Louis Bar syndrome) chromosomal ƒƒ In conjugation, the responsible plasmid is termed ‘sex
R

breaks + nonrandom rearrangements are seen. factor’ or ‘Fertility (F) factor’. F factor is actually an
episome.

4.7 BACTERIAL GENETICS Methods of gene delivery

ƒƒ Transfection is the process of deliberately introducing
ƒƒ Genetic material in bacteria: Genetic material in
nucleic acids into cells.
bacteria are portion of chromosomal DNA (plasmid) or
ƒƒ There are many methods of introducing foreign DNA
extra-chromosomal elements (episomes).
(gene) into a eukaryotic cell :
°° Episome is an accessory Extrachromosomal replicating
genetic element that can exist either autonomously or °° Electroporation
integrated in a chromosome. °° Magnetofection
°° Sonoporation
°° A plasmiD is a small, circular, ds DNA molecule, which
is distinct from chromosomal DNA. It is physically °° Gene gum
separate from DNA (i.e. and cannot be integrated °° Dendrimers etc.
like episome). It can replicate independently of, °° Site directed recombination
218 chromosomal DNA within a cell.

High-yield Points
44 Transferable drug resistance is seen in Enterobacteriaceae,
vibrio, pseudomonas, pasteurella.
44 Transposons are called jumping gene.
44 Plasmids are circular DNA molecules present in the cytoplasm of
bacteria, capable of autonomous replication.
44 Bacteriophage are viruses that parasitize bacteria and consist of
a nucleic acid core and a protein coat.
44 The plasmids determining penicillin resistance in staphylococci is
mediated by transduction.
44 Largest fragment of DNA can be cloned in --- cosmid
Lactose Operon /Lac Operon
ƒƒ Lac operon is cluster of genes encoding for proteins
involved in lactose metabolism in prokaryotes.
ƒƒ RE restricts viral (bacteriophage) replication.
ƒƒ Cuts at palindromic site. G
ƒƒ Methylation of host DNA (Bacterial DNA) protects from E
RE. N
ƒƒ RE type 1& 3 require ATP & have both endonuclease & E
methylase activity. Type 2 does not require ATP. T
I
Cloning C
S
ƒƒ Multiple copies of DNA are produced using vector.
ƒƒ Steps: Fragmentation → ligation → transfection →
screening of successfully transfected cells.
ƒƒ Classical example : Myeloma cells were fused with B-cells
to form hybridoma cells in presence of PEG (Polyethylene
glycol) → Grown in HAT (Hypoxanthine aminopterin
thymidine) medium → Used to produce monoclonal
monospecific antibodies (mabs like herceptin).

/e
ƒƒ Operons and polycistronic mRNAs are common in bacteria ƒƒ Cloning vectors & their capacity:-
but differ in eukaryotes.
ƒƒ Genes in Lac operon: Vector DNA insert Remark

14
Lac Z : codes for b galactosidase size
Lac Y : codes for permease •• Plasmid 0.01- 10 kb Smallest, m/c used
Lac A : codes for thiogalactoside transacetylase
•• Lambda phages 10-20 kb Confer antibiotic
ƒƒ Regulatory portion in Lac operon: resistance
Positive regulator : CAP protein & cAMP
S,
•• Cosmid 35-50 kb -
Negative regulator : Repressor protein
Promotor (P) region : Where RNA pol binds •• BAC 50- 250 -
Operator(O) region : Where repressor protein binds. •• YAC 500 - 3000 Largest
AM

4.8 RECOMBINANT DNA TECHNOLOGY RFLP

ƒƒ DNA is cleaved into fragments using restriction enzyme.

O

Recombinant DNA Technology ƒƒ 2 Types:

ƒƒ Joining of 2 different DNA via enzyme ligase. RFLP
ƒƒ cDNA is used to insert in to the vector DNA. cDNA
R

(complimentary DNA) is the double stranded copy of the

cellular mRNA.
ƒƒ To produce insulin in bacteria for therapeutic purpose, Length polymorphism/ SNPs (Single nu-
initial material is mRNA from beta pancreatic cells of Tandem repeats cleotide
polymorphism)
human.
ƒƒ Genomic DNA library : is the complete genome of a
Microsatellite Minisatellite 90% cases
particular organism.
(Short sequence ( 1-3 kbp motif,
ƒƒ cDNA library : contain those DNA sequences that are
2-6 bp repeats usually 15-70 bp)
transcribed as mRNA. < 1kb)

Restriction Endonuclease
ƒƒ RFLP analyze single base changes (e.g. Sickle cell anemia)
ƒƒ Key enzyme used in recombinant DNA technology.
or deletions or insertion of DNA into a restriction fragment
ƒƒ Makes 2 incision through sugar phosphate backbone
(e.g. thalassemia).
(phosphodiester bond) of each strand of dsDNA called as
recognition sequence/ restriction site.
219
 ƒƒ Used to detect:- ƒƒ Clinical uses:
G 1. Prenatal diagnosis 1. Detection of infectious diseases
E 2. Trace chromosome from parents to offsprings 2. Detection of variation & mutation in genes
N 3. Linkage of polymorphism with gene mutation (Prenatal & postnatal d/g).
E 4. Direct detection of mutation causing d/s. 3. To detect allelic polymorphism
T 4. DNA fingerprinting
I
C High-yield Points
S 44 Vector used to increase the yield of protein produced in High-yield Points
recombinant protein synthesis → Inducible promoter region. 44 DNA sequencing is a process of determining the exact order of
44 IPTG is a compound added to cells to activate a promoter gene. 3 billion bases.
44 Chromosomal walking involves repeated cloning of overlapping 44 Gene mapping is localization of specific genes to distinct
DNA segments of 100-200 kb. chromosomes.
44 In patient with DNA mismatch repair deficiency mutations accu­ 44 Blue white screening is used for detection of successful ligation
mulate in microsatellite repeats, k/as microsatellite instability. & identify desired DNA into plasmid vector.

/e
DNA AMPLIFICATION TECHNIQUES
4.9 GENE THERAPY
ƒƒ Also called as NAAT (Nucleic acid amplification

14
techniques). For Genetic Diseases
ƒƒ Commonly used techniques are:-
ƒƒ ADA- SCID
1. PCR & Real time -PCR (RTPCR)
2. NASBA (Nucleic acid sequence based amplification) ƒƒ Chronic Granulomatus Disorder (CGD)
3. Ligase chain reaction ƒƒ Hemophilia
S,
4. Gap LCR ƒƒ Under trial for congenital blindness, lysosomal storage
disease and muscular dystrophy.
PCR (Polymerase Chain Reaction)
For Acquired Diseases
AM

ƒƒ Discovered by Karry Mullis.

ƒƒ Use of 2 oligonucleotide primer. Mutant & Wild type
oligonucleotides can be used as a probe.
ƒƒ Rapid automated method of amplification of specific DNA
sequences.
ƒƒ Enzymatic non recombinant DNA amplification.
O
ƒƒ Cancers : Ad.p53 for head and neck cancer, prostatic
cancer, pancreatic cancer
ƒƒ Neurodegenerative diseases such as Parkinson's Disease
and Huntington's Disease

ƒƒ Viral infections (e.g. influenza, HIV, hepatitis), heart

ƒƒ PCR requirements:
disease and diabetes.
°° Taq polymerase enzyme & SYBR green dyes.
R

°° Mg
++

°° d-NTP to facilitate amplification

°° Primers
4.10 POPULATION GENETICS
°° Thermus aquaticus bacteria, which has only DNA
polymerase activity. The Hardy–Weinberg Law
°° In RTPCR thermus thermophilous is used, which has ƒƒ States that allele and genotype frequencies in a population
both reverse transcriptase & DNA polymerase activity. will remain constant from generation to generation in the
{ N Dideoxynucleotides & probes are NOT required}Q absence of other evolutionary influences. Hardy–Weinberg
ƒƒ Steps: law fails if there is non-random mating, dynamic
Sample DNA population, small population, natural selection, genetic
↓ (denaturation) drift, gene flow, migration & meiotic drive.
ssDNA ƒƒ Hardy–Weinberg law is only applicable for :
↓ (Annealing) Random mating population,
Primer (ds DNA) Infinitely large population,
↓ (extension) Static population
220 Polymerization ƒƒ Punnett square for Hardy–Weinberg :
Multiple dsDNA
 Females GLOSSARY
G
A(p) B(q) Molecular Dogma in bioinformatics era E
A(p) A(p) AB(pq) ƒƒ Genome is total genetic component. All the DNA contained
N
Males

B (q) AB (qp) BB (q )2
ƒƒ Punnett square for Hardy–Weinberg : The sum of the
entries is p2 + 2pq + q2 = 1, as the genotype frequencies
for an infinite population size) must sum to one.
4.11 OTHER IMPORTANT POINTS
ƒƒ Single nucleotide polymorphism (SNP) transgenomic Wave
Denaturing High Performance Liquid Chromatography
(Wave /DHPLC) is an approach that can detect single base
in a cell.
E
T
ƒƒ Transcriptome is total mRNA produced by a cell.
I
ƒƒ Proteome is sum total of all proteins synthesized by a cell.
C
ƒƒ Chromosome: a grouping of coiled strands of DNA,
S
containing many genes.
ƒƒ DNA (deoxyribonucleic acid): a molecule found in cells of
organisms that encodes genetic information.
ƒƒ Gene: a biological unit that codes for distinct traits or
characteristics.
ƒƒ Genome: the complete set of genes in a cell.
ƒƒ Genotype: the genetic constitution of an organism.
ƒƒ Metabolome: Complete set of LMW (Low molecular

/e
pair differences b/w otherwise identical 750 bp fragments weight compounds in a cell at a given time.
of DNA.
Imp Disciplines

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ƒƒ BLAST (Basic Local Alignment Search Tool ) is used to
compare short sequences of proteins & nucleic acids. Specialty/ Definition/Study of
Discipline
ƒƒ Oligomisnorm gene used in primer → Error based PCR
Genomics deals with the discovery and noting of all
ƒƒ Luciferases are oxidative enzymes used in bioluminescence the sequences in the entire genome of a
in genetic engineering.
S,
particular organism.
ƒƒ CASPases are 'cysteinnyl aspartate specific proteases'.
Epigenomics Alteration of chromatin & histone proteins &
ƒƒ DNA estimation can be done by → Spectrophotometer. methylation of DNA sequences.
ƒƒ cDNA is preferred for the preparation of gene library.
AM

Proteomics Entire library of proteins & its relation with

ƒƒ Entrez Gene is Gene map database which includes genes
diseases. large-scale study of proteins,
that have been completely sequenced. particularly their structures and functions
Microbiomics Is the study of microbes interactions within an
High-yield Points ecosystem
O

44 Entrez Gene & Hap Map are used to identify single nucleotide Genomics deals with the discovery and noting of all
polymorphism that may contribute to pathological conditions the sequences in the entire genome of a
44 CDART, MMDB, and VAST are used to analyze the domain particular organism.
R

structure & 3-d structure of proteins Metabolomics Aims at determining a sample’s profile of
LMW compounds at a specified time under
specific environmental conditions.

[IMP. SYNDROMES ARE GIVEN IN SYNDROME

SECTION]

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 Exam Oriented Prototype IBQs (Including PGMEE 2016-17 IBQs)
G
1. Karyotype of a cytogenetic analysis is shown below. 3. Analyze the following pedigree and give the mode of
E Diagnosis is:-  inheritance:
N
E
T
I
C
S

a. Autosomal recessive
b. Autosomal dominant

/e
c. Mitochondrial inheritance
a. Downs syndrome d. X linked dominant
b. Turner syndrome

14
c. Edward syndrome 4. The structure of RNA molecule has been shown here,
d. Klinefelter syndrome it represents:- 

2. Which of the following disease corresponds to the

pedigree chart given below:
S,
AM
O

Ans.
1. b
2. d
R

3. c
4. b

a. Achondroplasia
b. Wilson’s disease
c. Hemophilia.
d. MELAS

a. mRNA
b. tRNA
c. rRNA
d. None of the above

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