Molecular

Basis of

Inheritance
 Introduction:

DNA-genetic organisms.
material
for most

controls
the inheritance trails
-

of
from to next.
the
one
generation

NUCLEIC ACID

RNA
I

DNA

Ribonucleic
Deoxyribonucleic acid.
acid

↓
↓
material
Genetic
material
Genetic in riboviruses.

all
in
organisms d
except some
functions as

viruses.
A
messenger carrying
information
genetic
A
adapter for picking
up amino
acids,
structural
and
catalytic
molecules in some

cases.
 DNA-Deoxyribonuclei
acid:

-> substance
Acidic

->
nucleus.
present
in

Friedrich Meischer
firstidentified by
->

1869.
in

L I

↳
called it NUCLEIN

S L ALTMANN, -found substan

these -

-
ces to be acidic in nature

> NUCLEIC ACID.

Length
of
DNA -number
of nucleotides

nucleotides-Base pain.
pair
of
(bp)

CHARACTERISTIC
->
OF AN

ORGANISM.

Examples:
 circular
1.
In bacteriophage-SSDNA,
5386 bases.

Lambda(x) Linear
2.
phage-dSDNA,
48502bp.

3. Escherichia coli
dSDNA, circular

4.6x100bp

4. Human linear
genome ASDNA,

haploid 109bp
↳
3.3 x

2x3.3x109 6.6x109bp
Diploid
-> =

STRUCTURE OF POLYNUCLEOTIDE CHAIN:

DNA-basic unit -
nucleotide

NITROGENOUS PENTOSE PHOSPHATE

BASE
SUGAR GROUD.

↓
↓
Two Deargeibose
types: (C5 1004)
1. Purines:
-
DNA-
 Ribose
9- membered

double [C5 +1005]

-
eing NA.
&

Structure

with N-1,3, 7 and 9.

EXAMPLES -

Adenine, Guanine
(A) (q)

2.
Pyrimidines:
Heterocyclic, 6-membered, single ring
structure N-1,3 position
with

Example i

Cytosine, Thymine Vracil

CC) (T) CUS
- ↓ ↓
Both DNA+ DNA RNA
RNA
 Linkage
↓

polynucleotide
has
chain
linking
components
among

PHOSPHOESTER
N-GLYCO SIDIC

LINKAGE: LINKAGE:

of
Between OH when
of
I'c 5'
of pentose of nucleoside

I
and
sugan
nite
presphate
genous group.
base. (C- 1 of sugar)
I -(position
↓
NUCLEOTIDE
purines -> 1-9
A(G) is
 I
geosidic
einbiage l'of of sugar
->
I'-position
Pyrimidine- of T/C.

linkage
-
Glycosidic linkage
↑-

forms
a NUCLEOSIDE.

Nucleotide +

Nucleotide

to
5
-

linked 3
through
HOSPHODIESTER
↑
LINKAGE

form
to DINUCLEOTIDE
 Every polymer-I. face phosphate
at 5' end

-
This
is the send

of polynucleotide
chair.

3'C
2.
Free OH
of group
↓
-' end chain.
of
polynucleotide
BACKBONE chain
of polynucleotide
↓
Sugar Phosphates
-

Niteo linked to
base
sugar
->

genous
projects
from
it.
 DERIVATION OF DNA STRUCTURE:

Two lines
of investigations helped in

denivation
the
of DNA structure:

a)
X-ray Crystallography:
Maurice
Wilkins and Rosalind

Frambein-obtained
very fine
pictures
DNA.
X-Ray diffraction of

suggested structure
of DNA was

sort helix 3.
with 4n
of
a

periodicity.
 But model
peroposed.
was
no
definitive

b) Erwin Rules:
Chargaff's
Chargaff" along with
colleagues

base
performed competition
studies

certain
and
put forward generalisati-
DNA
for
-
ons double stranded

Chargoff's rule

=> Not
applicable for single-stranded
DNA.

1. Purines occur
and pyrimidines
 in
equal amount.

2. Purines -> A +

Pyrimidines -> C+ T

A G c T.
+
:
+
=

= [This
3.
1 value constant for
a
all
species. I

4. Base ratio AtT-

specific
for
c G
+

a
species.
used to
species.
->

identify
one-probaryotes
·Less than

Ex: E.coli =
0.92

one--Eukaryotes
·
More than

Ex:Humans-1.52

5. and
Sugar
deorgribose phosphates
residues occur
equal
in number.

with
6. Purine adenine isequimolar
=T
thymine.
A

with
Punine
guanine equimolar
7. is

GE C
pyrimidine cytosine.
 Watson and Crick
Francis
James

proposed douber helic model

for
steructure DNA
of

Major hallmarks was their

proposition
-

pairing
was base blue the stands
two

chains.
of polynucleotide

-man
by
observations
the Fair

chargaff.

since
they
->

complementary
are to

one another, if you

know the

bases one strand,

sequence of your
in

find other
can out the stand.

A
= T

G C
=

G T AATCCTG

!++ad
Each strand also act
as a
template
 strand DNA.
for new
the
of
Questions:

1.
CTAGGCAGC
-

GATCCGT CG

2. AGUCCAGUA- UGAGGUCAU
Features:

DNA consists
of
. two

chains
polynucleotide
Backbone
sugar
-
-

phosphate + bases

inside.

2. Two chains awn in

anti
parallel fashion
5'- 3'
with
polarity
one
in and 3'-5'

polarity in other.

3. Bases
through
paired
Hydrogen
bonds.

A
T;G C.
=

purine
always opposite
*

to a
pyrimidine
4. Double chain-

twisted a
in
right-handed
helically
fashion.
5. Each
of double belive
or
pitch
tur n

is
culine 3.4mm.
Roughly 10
bp
of
the

between
Distance
in each
pitch.
two
adjacent base
pains -0.34 nm.

6. The stacks
plane of one bass
pair
over the other in a double helice.

This H-bonds-confin
along with

to helical structure.
stability
the

CENTRAL
DOGMA of Molecular
Biology:
->

explains-one way or unidirectional

flow of information
master
from
Copy
DNA
to
copy
working
RNA
and

RNA
to molecule
from building
or

trait molecule
expressing polypeptide.
Proposed by Francis Crick.

Replication

⑮ A
transcription mRNA
translation
>Protein

REVERSE CENTRAL
DOGMA/ TEMINISM

- >

Exception
viruses.
->
Only some
in
 H. Temin and D. Baltimore
->
Given by (1970)

The viruses
produce
->

enzyme
an

called
TRANSCRIPTASE.
This
synthesizes DNA over RNA

template.

IMPORTANT in
understanding
- >

cancer cells.

~
-
This Nobel Price.
given a

8
TRANSCRIPTION
TRANSLATION
I
D ·A RNA > Protein
W
REVERSE
TRANSCRIPTION

-NGHELIX:
-> Distance blw two consecutive

base min-0.34x10m.
pains=0.34
-> No.
up
humans
in
-
6.6x109bp.
 human DNA=6.6x109x
length
->
of
0.34 x 10-i

=2.2m.
More than
lengths
of atypical
nucleus (10 m).

-> Number inEcoli

of base
pairs
= 4.6x104

4.6x10 10-9
-Length
of
DNA 0.34
=>
x
x

I 1.36 mm.

This lot
larger
isa than 1
m
is
which the cell
size

Thus DNA
only
can be accommodated

packaging
in small
a area
through
compaction.
or

DNA
packaging Probaryotes:
in

the cell
DNA is not scattered
throughout
->

don't well-defind
though they
even have a
 nucleus.
SUPER CONED
DNA
->
-

found cytoplasm-INSTAGE-
in

->
coils maintained non-histone
by
basic POLYAMINES
protein which
have

change.
positive
Nucleoid
Packaged of DNA-
structure
or

Genophore.

Packaging
DNA in
Eukaryotes:
Organisation comple
-> - much more

out
carried
and
by a set
of
positively changed basic called
proteins
HISTONES.

rich basic
in
amino acid

residues-lysines and

arginines
with
charged
side chains.

->
Five
types of histone
proteins:
H;MeA;MzB;Me and
My

↓
4 Ma, My and

them
of -> MA, ty
 ↓
pairs form
to
occur in

HISTONE OCTAMER on

mu-body.

charged
Negatively
DNA-
wrapped
histone
around the
charged
positively
octamer to structure
form
a

-> NUCLEOSOME-

H2A
↑Ha
↑
Hy
H3

Around
200bp of DNA-
wrapped
over me
body
to
complete
about

11 twes.
 100 60h.
x

of
size nucleosome ->

DNA
present
between
two adjacent
nucleosomes - LINKER DNA

(with about
80bp.)

Nucleosome has unit

repeating of
structure in the nucleus called

CHROMATIN

-> The beads

nucleosome-
sering
on a

appearance.

further
coils - SOLENOID/
CHROMATIN
FIB RE

(Diameter-30nm).

condensed coiled
further
metaphase
at stage
all division to
of
form chromosomes.
 chromation at
>Passaging of
level additional
higher an
requires
called
set
of proteins
NON HISTONAL CHROMOSOMAL

(NAC) PROTEINS.

1.
scaffold Functional Regulatory
or
2. 3.

structural NHC NHC

protein
Ex: DNA Ex:
NHC HMG
polymerase, Chigh
RNA
polymerase. mobility
group
proteins
contro
that

gente
expression.

Chromatin on

staining
the basis
of

1. HETERO CHROMATIN 2. EUCHROMATIN

stained stained
darkly lightly
* *

region. region.
Loosely packed
*
is
Chromation
*
 chromatin
densely packed
Transcriptionally:
* Transcriptionally
*
-
active.
inactive.

e
Chemical
composition
of chromosome:

DNA =40%

RNA 1.2%
=

50%
proteins
Histone =

Acidic 8.5%
proteins
=

Lipid Traces
-

+ 2
Fett
+2
Ca
ig
-
Traces.

THE
SEARCH FOR
GENETIC MATERIAL:

that
Experiments
proving DNA-genetic
material:

1. TRANSFORMING PRINCIPLE:
m e re

Experiment conducted
by
FREDERICK

GRIFFITH 1928. evidence.

great
-

in

He selected strains
of bacterium
two

Streptococcus pneumonial
/Pneumococcus)

S-111 R-11

S -
 ->
smooth/ ->

paugen/
type
capsulated non-capsular
type
Ted

have
-

a mucous
to
no mucous

/polysaccharide) and
coat

coat and produce

rough
produce smooth colonies

colonies
shiny
in culture
plates
b
non-virulent
↓ and do
not

virulent
and
cause

cause
pneumonia. pneumonia.

conclusion:

1.R-strain bacteria had somehow been tran-

 the killed S-strain
sformed by heat
-

basterior.

2. occured
This
perhaps due to
absorption

of
some
transforming principle on

bacteria
substance
by rough type from
heat-killed bacteria.
smooth

3. This
enabled the to
R-strain
synthesize
and
a smooth
polysaccharide
coat

become virulent.

This
->

due to the
be
must
transfer of
material.
genetic
Deawback: Biochemical of genetic
nature

material-not
defined from
his
experiments.

BIOCHEMICAL CHARACTERISATION TRANSFOR-

of
-

MATION PRINCIPLE:

Oswald
wwo-? 1.
Avery
2. Colin Macleod

(1944)
3.
Maslyn Mocanty
b
 in-vitro to
experiment
repeated
biochemical nature
of
identify
the

transforming
substance.

↓
They proved substance
this
that - DNA.

Prior
to this, material
genetic thought
-

be
to
protein.

what
did
do?
they
1.
They purified
biochemical i.e.

DNA, RNA heat

proteins, from
scells
killed to see which over

could
transform live R-cells into

S-cells.

They
discovered alone
2. DNA
from
S-bacteria caused R-bacteria
to

become
transformed.
They discovered that
protein
-
5. also

digesting enzymes
i.e.
protease and

is. RNase did

RNA-digesting enzymes
 transformation
affect
-

not

not
substance
Transforming
... was

or RNA.
a
protein

4.
Digestion DNase
with did inhibit

transformation
↳
that the
suggesting
DNA caused

transformation.

many biologists
Drawback -
not were

convinced.

EVIDENCE EXPERIMENTS
2.
from with

BACTERIO PHAGE:

who? Alfred and Marina

 Chase
(1952)

Why? unequivocal proof-DNA

material.
genetic
-

How ?

1.
They worked a virus
with

↓
i
bacteriophage.

infects
bacterium Escherichia coli

and inside
multiplies it.

Tz made
phage up of-DNA+
-

protein
coat-

:Most suitable material

to determine

Whether
DNA contains
or
protein
the
for
information production
virus
of new
particles.

Functions
of DNA and could
proteins be

them with
by labelling
out
found
tracers.
radioactive

DNA
contains
phosphorus
but not
sulphur.
...
phage DNA-labelled p
with
 How?

to
by growing bacteria
infected
with in medium
culture Poy
phages

Another contains
protein
of phage
-

sulphur but no
phosphorus.

·Phage protein coat

was labelled with

by
5
growing
bacteria
infected
with

another medium
culture
phages
in

containing "Soy

labelling,
3
After steps were
followed:

Both
Infections:
1.
labelled
nee
types of phages
were allowed to
infect
normally cultured bacteria

in experiments
separate

BLENDING:Bacterial
cells
2.
- e
were
agitated
ina blender to break the

contact between virus and

bactired.
 particles
were
The
3.
GATN: virus

by spinning
bacteria
from
the
separated
them a
in
centrifuge.

centrifugation-bacterial
cells
After
showed the
presence of radioactive

DNA labelled with

P32

while,

radioactive labelled with

protein
the bacterial cells
appeared outside

i.e. in medium.

Labelled DNA- also in the

found
was

next
of phage.
generation

viruses
Bacteria-infected
with

that had radioactive

proteins
were not radioactive

that
indicated
This proteins
didnot enter the bacteria
 the
from viruses.

DNA- material
genetic
.

to bacteria.
passed from virus

PROPERTIES OF GENETIC MATERIAL:

A molecule that can act as

genetic
material
fulfil following
must the

criteria:

be stable.
structurally
and
Chemically
.

c. should be able to
generate
its
replica
 <replication).
slow
3. Should
provide
the
scope for

mutation evolution.
required for
4. Should be able to
express itself the
in

form of Mendelian characters.

Why DNA-preffered genetic

material?

1 Free L'OH RNA-makes it more

of
labile
degradable.
easily
and

DNA in
comparison more

Stable.

of thymine(5-Methyeuracil)
2. presence

at
place of Uracil
the
confers
stability to
additional DNA.

3. RNA-unstable, mutates
faster.
consequently
viruses
having
RNA
directly code
can
genome
for of
synthesis protein.
can
easily
express characters.
 -world:

RNA-firstgenetic material.
·

as
RNA-acts
genetic
·
as material well as

catalyst.
a

importantbiochemical reactions
·
Some

that RNA
catalysed
by catalyst
-

are

ribozyme and not

by protein
enzymes.
Example:
Ribonuclease P-
derage
Smurps speicing
-

Peptidyl transferase
band
-

Peptide
formation.

But, RNA and

reactive
catalyst
-
as a

unstable.

DNAevolved
from RNA chemical
with
.

it
making
stable
modifications
more

DNA
being
double stranded
and
having
strands
complementary further resists
 changes by process of repair.
a

evolving
molecule
structural
RNA-adaptin
and in some eases - CATALYTIC.

RNA DNA
function
and as
genetic
. can

material, DNA-more
but stable

material.
preferred for storage of genetic
For transmission
of genetic material

to
RNA is better.

EPLICATION:
R
nee

proposed
a
Watson and Crick

DNA
scheme
for
the
replication. While
peroposing
double helical
of
structure DNA.

Process:Two DNA stands

separate
↓

act as
for
template synthesis
strands.
of
new

-
 After completion
of replication
↓
Each DNA I strand
parent
I
newly synthesi-
strand.
yed
-

This
scheme semiconservatives DNA

replication

Experimental peroof:
-

1. Matthew Meselson and Franklin

Stall

(1958)
performed following
experiment heavy nitrogen
using
(IN) E.coli.
in

rocess.
-

E.coliin
a contain
medium
greue
15

Nyc the
-ing only mitogen
as source

for many generations.

"N-heavy of nitrogen
isotope
 was
incorporated newly
into

DNA well as
synthesized as

others
Nitrogen containing
compounds.

distinguished
DNA
molecule
heavy
-

normal
from the
by centrifugation
DNA

cesium chloride (CSC)

gradient.
density
in

A dense solution CSC

of on
centrifugation
forms density
gradient of
bands a

solution
of lower
density
at
top
the

increases
that
gradually
towards

the bottom
with
density.
highest

~I

Cells medium
were
transferred into a

normal
with "NHyc
↓

various
samples were taken at

intervals
time
definite as the

cells multiplied, and cictracted

the DNA remained

that as

double-stranded helix.
 ↓
The various samples were
separated
independently
on
ascIgradients
to

the
measure densities of DNA.

↓
:the extracted
DNA the
from
culture (
after generation
i.e. minutes has
after
20

hybrid
or intermediate
density.
↓
from
DNA extracted the

culture
after
another
generations
2nd
i.e.
generation or 40
after
minutes equal amounts
of
(NN's) and of
hybrid
DNA

(N'YN'Y.
-

light DNA

Increase the amount

in
of
light
DNA and decrease in

DNA
hyberid amount
can be

possible due semiconservative

to

mode
of replication.
 proved
2. et.al.
Taylor semiconser
-

-
vative
mode of chromosome
replication
eukaryotes using
in
thymidine
teitiated

(A-thymidine thein root

of
Vicia

beaus)
faba (Faba
 semiconservative
3. Cairns -

peroved
-
E.coli

of replication
mode in

titiated
by using thymidine
C +dR)
autoradiography
in

experiment.
He also o-model
proposed
circular
DNA.
for replication in

Machinery Enzymes/DNA
The and the

Replication
Mechanism:

->
process
of replication requires a

set
of enzymes.
Main
enzyme DNA-dependent DNA

polymerase.
-
 with
highly efficient 2000bp
ability to polymerise some

per
second.

->
Not
only polymerases
these have

to
be but
fast
to
also
they
have

catalyse higher
with
degree of
accuracy
as mistakes
any ↓
MUTATIONS.

whole
genome of
Ecoli 4.6x10p
having
within 38 minutes
replicated

Steps of
DNA
replication:
1.
ORIGIN REPLICATION:
OF

Replication
begins
at
particular
-

called ORIGIN OF
region
&
EPLICATION.

to OR1-
provide
->
we need

vectors.

prokaryotes
-> ONE ORI

Eukaryotes many
 Ecoli-ori-c
·

In
-

2. Activation
of deoxyribonucleotides:

Four
types dearcyribonucleotides.
-

3
d AMD dATD

d GMP 2 > dG TD
t
HPOy
d TMP dT TO

d CMP d CTP

deoxyribonucleotides
activated
by
... are

phosphate, energy and enzyme

phosphorylase triphosphate
into

state.

Dearcyribonucleosidecaniphosphate

I serve as a 2. Provide

substrate energy for

polymerisation
as
reaction
 two terminal
phosphates
a
in
deorcynucleoside
triphosphates high
-

energy phosphates same -

as ATP.

mming
of helice:

Unwinding
of double helical
parental
·

molecule about
brought by
HELICASE
enzyme is
which ATP
dependent

of
Unwinding
molecule into
·
DNA

two strands
7
results in a
of
formation
Y-shaped structure.
↓
called
Replication
fork

E
single
stands stable
with the

help of strand
single proteins
binding
SS Bi
-
 51 31

3 51
T
CONTINUOUS DISCONTINUOUS
-
SYNTHESIS 5YNTHESIS

-
Newly
3 51
synthesized
- 4
5
31
strands

REPLICATING FORK.

unwinding coiling
Due to
super
a

end
developed
gets on the
of DNA

to
opposite replicating fork

Unwinding
done ENZYTE
by
TOPO ISOMERASE.

In
prokaryotes has
DNA
gyrase
topoisomerase activity.

4
meN
PRIMER STRAND: OF

New stand has to

synthesized
->
be

parental
strand.
opposite
 III
-> DNA
polymerase
true
explicate
in E.coliincapable of initiating
DNA unable to
synthesis it is

deposit the first nucleotide in

daughter.
strand.

... An PRIMASE
enzyme
↓

synthesizes a short
primer
strand RNA.
of

This strand
Acts
primer
as a

stipping
to
stone start

errorless
replication.

Once DNA
synthesis completed
RNA strand
is then
:primer
removed
enzymatically.

5
meNADS:
NEW
OF

polymerase
DNA

dependent
DNA

in
catalyse polymenation only
 5'- 3'
one direction:

...
Replication continuous
is
on

template strand
with
polarity
-
3 5'

↓
This
is DAUGHTER
LEADING
STRAND.

Replication discontinuous
is
on

the other with

strand
polarity
5'-3' and it in
happens
fragments
called-OKAZAK1

FRAGMENTS.
-
These
E
fragments later

by
joined ligase.
DNA

* In
eukaryotes
--

Replication of DNA

takes
place at
s-phase.
divisions
Replication
of DNA+ cell

cycle
↓
coordinated.
 A in cell division results

failure
POLYPLOIDY.
in

DNA
polymerase stypes:

Exonuclease
Polymerase
activity activity
-
I 5 3 5
-
3

↓
(aub [removal
eng
adds
enzyme) of RNA
primer 1000

and 7 T
=

nucleotictes

Cthy
mine
peer
minute.

dimen

formed by
unrays]
and 3'- 5'

5 3
-
:
-

3 5

Rate
-> 50

nucleotides/
minute.
 -
5 5'- 3' with
: 3

polymerisation
rate
of 2000bp
second.
per

Synthesis leading/continues
->
strand
of
is
quick and one
requires primer.

discon
of logging stand
->
synthesis -

- tinuous and
many
requires primers.
DNA
polymerase III
-

polymerizing
Main
enzyme

TRANSCRIPTION
nee

process
genetic information
of copying
into RNA-
from of
one strand DNA

TRANSCRIPTION

-> Like DNA

replication, principle of
complementarity
governs process
the

adenosine
of transcription,
but

forms
complement base with
pair
uracil instead
of thymine.

But,
- DNA
In DNA
replication total
of
organism
gets
duplicated

Transcription onlysegment
a

of and
DNA
only
ONE strands is
copied
into
RNA: one

only
strand transcription
in is

template
stand.
Reasons

↳ -

strands act two RNA

If both

molecules
as
template if
-
RNA produced
code for
molecules with
simultaneously
 would
different
be
sequences.
↓ complementary
other
to each
they
Then when

code
for proteins, ...
would
form
-
a double-
proteins
different
- Stranded RNA.

amino ↓
different
acids. would
This

.. the
one
segment of prevent
codes
DNA --
for translation
of
into
two
different RNA

proteins. protein.
:This
complicates
genetic
the

information
transfer machinery.

TRANSCRIPTION UNIT

to

that
segment of DNA
takes
part transcription
in

TERMINATOR
STRUCTURAL
PROMOTER
GENE
 strand
Template
stand
and
coding
DNA
dependent RNA
polymerase
in
catalyses polymerisation
the

5'-3'
just one i.e.
direction
polarity

Transcription
start

Site

PROMOTER Non-coding strand) TERMINATOR

strand
3 Template 5/
structural
gene
5) 3
stand
coding

The strand
that has the
polarity
3- 5' acts
as a
template/non-
strand/anti-sense stand.
coding
stand
other 5'- 3'
with
polacity
RNA
has the same as
sequence
except thymine at the
place of
during
uracil: It
is
displaced
transcription. This
strand

strand/sense strand/ non-

coding
template
strand.
 Transcription
start

Site

PROMOTER Non-coding strand) TERMINATOR

strand
3 Template 5/
structural
gene
5) 3
stand
54 3,
coding
Promoter is
upstream
present
-

5'
towards
of structural
end
gene
of transcription unit -
In

polarity
of coding
to
reference
stand.

binding
provides
DNA
sequence
site
for
RNA
polymerase.
Presence
of promoter transcription a
in

unit
template and coding
defines
strand.
28 promoter
with termination
changes position
definition of template and coding'
strand reversed.

Binding
sites RNA
for polymerase
lie within
promoter sequence.
Certain
short
sequences within
promo
-
Heer Sites conserved and called

RECOGNITION SEQUENCE.

Terminator 3'end
at
present
of codingof
strand
defines
-

the end
process of transcription

Meet
gene

unit in e
inheritance.

Genes located DNA.

-
-
on

tRNA
for
DNA or
sequence
coding
-

rRNA
molecule-defines a
gene.
CISTRON-functional
unit
-

of gene.

>
Segmentof DNA
for
coding
a

polypeptide.

STRUCTURAL GENE IN

TRANSCRIPTION UNIT
 MONOCISTRONIC POLYCISTRONK

eukaryotes prokaryotes
(mostly or bacteria

mostly.
X X

synthesizes only synthesizes

-
-

type different
one
of
on
polypeptide
or
proteins
protein. polypeptides.

They
-

have
↑

interrupted
sequences
coding
il in
genes
eukaryotes
are

split.

sequence
Coding on
expressed sequences
-

called EXONS

appear mature
in or

processed
RNA.

Exons-interrupted
by
 I

intervening
sequence that
dont

nature
appear in on
processed
RNA.

Split gene arrangement further

definition of gene
the
complicates
terms DNA
in
of segment.

Splitgenes-discovered by:
Richard J. Roberts
philip
and

sharp.

TYPES OF RNA:

nee

mRNA ERNA tRNA

/Messenger RNA) [ribosomal RNA) <transfer RNA)

 &0% · 15%
5% total ·
·
of
RNA

smaller smallest
·
Longest . .

·
called ·
has structural ·
It is

templatel (forms
ribo soluble on

nuclear -

some) and
adapter
RNA and
messenger Catalytic
or role

informational during carries

RNA as it translation. amino

genetic
carries acids.

information
by
provided
DNA-

All are
three needed to
synthesize
ina cell.
protein

meeTION
IN PROKARYOTES

-> occurs CYTOPLASH

help
in with the

n e e
of

transcripting
enzyme.
-
DNA dependent RNA
polymerase -

only
is
 transcribe
of one
type
and all
types of
RNA

RNAs-RNA,
all three required
to

ina cell.
protein
synthesize
functional
completely
~ enzyme
catalytic
active
RNA
polymerase -

holoenzyme
e

made
up of polypeptides
(C2 BBN) =

- Enzyme without subunit

referred
to core
as
enzyme.
Three
steps:

!ON:
 or initiation
-

catalysed by
I
factor
factor.
to
binds the
promoter
site
of DNA
-

and
confers specificity.

If transcription
them
father-absent
a

specifically by
Starts care
now
enzyme
at base on DNA.
any

ATION:
2.

(core
- RNA
polymerase enzyme) -

the
only capable of catalysing
&

process of elongation.
3. TERMINATION:
-

Rho
factor (p) required for
-

ermination.

RNA
Polymerase
-

to
binds
promoter
the
of
region the
-

DNA and the

process of transcription
begins.
substrate
-

uses nucleoside triphosphates

as substrate

a
polymerises in
template
depended
fashion rule
following of complemen
-tarity.

Also
helps the
opening
-

in

of
elongation
helie and continues

Only a stretch
strant
of
RNA

attached to the
is
enzyme.

Once termination
polymerase reaches

both nascent RNA and

region
RNA
polymerase falls off
and it

termination
results
in of
transcription.

... Bacterial
transcription:
any processing
"mRNA does not
require
to become active.
 2.
Transcription
and translation

take
place
the same
compact
-

in

-went

translation
Many
3. times can
begin
mRNA transer
before fully
much the is

bed.
-

Transcription
and translation can

be bacteria.
coupled in

MeTION
IN EUKARYOTES:

Types enzymes:
transcripting
- -

of
IRNA in nucleus
polymerases
RNA
polymerase organelles.
2. in

Functions:

* RNA 5.85, 185, 28S rRNA

polymerase
I ->

Synthesis.

A RNA I -> hRNA

polymerase
(retrogenous
nuclear RNA).
* tRNA, ScRNA
RNA
polymerase I -

Ismall RNA),
conditional

5S rRNA -SMRNA
<small nuclear RNA

Nascent
RNA RNA
synthesized by
polymerase II is called
NA on

primary transcript.
↓
This
contains
both unwanted

sequences (introns)
base and

wanted base (eccous)

sequences

CAPPING
of
Addition
↓ ↳ ademplate
resides
done
addition
by
-
RNA SPLICING
-
Dozenbeiein
of methyl
GTP to 5'

end -

Catalyst
Guanyly
transferase.
RNA (mRNA)
MESSENGER
 -steps:

:Modification
of
5'end
by capping:
->

capping
at5' end occurs
rapidly
after
start
of transcription.
An unusual
nucleotide-Methyl
added
triphosphate
guanosine
is to

the 5' end huRNA.

of
7

This
process by
catalysed Guaryly
transferase.

why cap important?

is

essential
for formation
mRNA-
of
-

ribosome complex.
Translation
not
possible if
cap is
-

18SrRNA
lacking as
cap identified by
is

of ribosome unit

small

regulatory
RNA.

2.
mmeePOLYADENYLATION:
 -

Tailing-addition
of adenylate
residues about 200-300 at 'end

dependent
in
template
in manner

hmRNA with the

on
newly formed help

Poly
A
of polymerase.

3. SPLICING:
nee

removal interious
of
and
of
-

process

joining
in order.
of defined
exons a

->
Introns removed small nuclear
by
are

RNACSMRNA) and
complete
protein
called small nuclear ribonucleoproteins
or SURNPs (snumps).

->

Fully
is called
processed mRNA now

and
mRNA
transported of
out

nucleus translation.
for
SPLIT
gene arrangements represent
an ancient
feature
of genome.
Presence introns-reminiscent of
of
antiquity.
 splicing
the
Process
of
-

represents
of
dominance

RNA world.

GENETIC CODE:
nee

DNA/RNA-
carry genetic information
RNA-
expressed the
in
form of
made
proteins up of
20
different
acids.
amino

Genetic code blu

inter-relationship
is

nucleotide
sequence of DNA or mRNA
and acid in
amino
sequence a

polypeptide.
containing
It mRNA coded
an
is
sequence
for
information one acid
amino and

consists three nucleotides.

of

History:
GEORGE GAMOW-Physicist:
coined code.
genetic
-

argued that order

in to code for
 the code should
all 20 amino acids,
nucleotides.
be made
up of 3

BOLD PROPOSITION combination

as
of
↳x4x4 would by
generate
codons
generatingmore
-

codons than
required.

Importantdiscovery
-was the

result
by
of experiments
Marshal Heinrich
Niremberg
w. and 5.

Matthai later
by G. know
and

and.

Mathai
Nirenberg
and -
used

poly
synthetic URNA and
deciphered
the code this
translating
as
by
polyphenylalanine.
Hau Gobind Knorana RNA
synthesized
-

molecules with
defined combination
of
bases
(homopolymers and
expolymers.

Using
synthetic
DNA- he
prepared
 repeating
known
with
polynucleotide
Ex: CUCUUCUCUCr
Sequence
- -

produced
This
only
two amino acids

leucine serine

(cue) (UCU)

Severo ochoa
dedide enzyme is

xylase-helpful in polymer
rising defined sequences
RNA with

in
template independent manner

i.e. RNA.
of
synthesis
enzymatic
 Ser-serine

Phe-phenylalanine
Lew-Leucive

-
Ile-Isoleucine

Met -

Methionine

Val-valine

Ala-Alanine

The-inreonine

Pro-Proline

Tyr-Tyrosine
His-Histidine
cys-cysteine
Gen-Glutamine Tep
-

Tuyptophan
Asparagine Aeg-Arginine
Asm-

Glu-Glutamic acid
Gly-Glycine
 code:
Features
of genetic
1. TRIPLET CODE -
Each codon made
of
a re
we

there
adjacent nitrogenous bases.
acids
6)-code
for amino

3-
stop codons-do code
for
not

acid.
any amino

puntuations.
NATURE no
2. COMMALESS -

m e e te

BineR
OF
CODE
Same
inthe
degenerate.
codons: code is

Ex:

3
serine

6 codons
Leucine

ginine
Au

3
Proline

valine
4 codons.

Glycine
Alaneno

Theomne
-

↳G-iotinnine 3
man
Exception
ene
-

generic
 b
amino acids

specified by their

single
corresponding
codon

4. UNIVERSAL CODE:code
nearly
is universal
nee
ror-codes
for phenyl-
-

alanine

all
in

organisms.
some have
exceptions been
found in

mitochondria and
protozoa.

CODONNAL:AUG-
5.

codes
for
methionins dual
functions
and was

methionine
codes
for
->

->
acts
as intiator codon.

6.
NAL:Polypeptide chain termin-

termination
signalled
ation
by
three
-

3
codons- UAA (ochere)
NONSENSE
UnG(amber) CODONS

UGA (opal) or

STOP CODONS.
MineRLAPPING
CODON: Each codon

does
overlap
not

independent
is and

the next codon.

US
GE AND
obe:
Example-sickle all anaemia.
-

to

point mutation-change of single

base
pain
in
gene for beta
globin
in the
that
chain results
change of
acid
glutamate
amino residue to

valine.

meete
adapter
molecule

Francis
existence
formulated
by
-

Crick.

-was also known as soluble RNA

(SRNA)

before genetic code was

postulated.
15% total
cellular
constitutes
of the
-
 RNA.

-> Crick
postulated:
* molecule
presence of
an
adapter
A that would

I would read 2. to
binds an

the code acid

amino

(specific).
* It
acts as an intermediate

molecule blu triplet code

of
mRNA
and acid
amino

chain.
sequence of polypeptide
A all RNAs have the same

basic
structure

(almost).

I over 60 tRNA.
types of

-> 3-D structure tRNA

of
INVERTED L-SHAPED ERNA

(given by
Kim and
Klug).
This is the actual
structure.
A structure tRNA
secondary
-

of
CLOVER LEAF
was to be
proposed m o re
e

Fee tRNAS:

I
have residue 5'end.
at
guanine
unpaired
2. has CCA
sequence
-

on

3'end.

3. acid
Amino attaches to send.

tRNA- to amino acid

specific specific

For initiation tRNA

specific
↳

INITIATOR tRNA.

tRNAs
for stop
->
No codons.

There are three

loops intRNA:

binding
1. Amino
acye synthetase
loop/DHU loop-Isloop
5' end
from
2.
Ribosomal with
loop
binding
 bases loop
1st
>
impaired
'end also called
from
-

TPC
loop.

3. Anticodon
loop with I
impaired
bases.

Out
of 7-3 bases act as an

for particular
anticodon a

triplet codon
present
mRNA.
on
 TRANSLATION
nee

refers
to
polymerisation
the
of
to
acids
amino
form a
polypeptide.
order and amino
sequence of
-

acids
by
defined the
sequence
bases the mRNA.
of in

cellular
factory
responsible
->

for synthesizing proteins

RIBO SOME.

* Ribosome consists structural

of
RNAs and about 80

different proteins.
 state:
inactive
A Ribosome In
-

two sub-units
Exists as

small
large subunit
sub-unit.

*
For
binding aminoSage
are in
of

P-Site A- site

(Amino
(Peptidyl acye)
Sitz)

when the small subunit

of
encounters
ribosome an mRNA,
the translation
process of
of mRNA to
begins.
proteins

STEPS:
e

tRNA
"ATION
OF on
 OF tRNA:

-G
Happens
->

presence of
inthe

AMINOACYL- ARNA
enzyme
/

(E)
synthetase I

acid (AA) bind

specific
amino

with ATD.

E,,
Mg2t
AA, +
ATD >
-
AA,-AMP-E,

complete
+ D Pi

AA, AMP-E,
complex with
reacts

tRNA.
Specific

-> Now is
amino
acid
transferred
to
tRNA.

-> As a
result, and AMD
enzyme
are liberated.

↓
This
iscalled AMINOACYLATION of
tRNA.
 tRNA,
AA,-AND-E, complex
+

↓
+ AMD +
AA,-tRNA, E,

2. FORMATION
OF POLYPEPTIDE CHAIN:

Three
steps:
1. CHAIN INITIATION:

nee

requires I initiation
factors
·

and
in
perobaryotes
9
eukaryotes.
in

smaller
Binding
(i) mRNA
of
with

submit ribosomes (305/40s)

of

30S sub-unit +mRNA

↳
30S-mRNA complex.

(In eukaryotes-formation
of

compe
40S-mRNA
 30S-mRNA tRNA
with
(ii) BINDING OF meth

memionine
non
formulated
->

is attached RNA
with in

with
eukaryotes and formulated
prokaryotes.
30S-mRNA complec +
tRNA meth

&
its
d GTP

30S -mRNA-tRNAfNeth

(iii) Attachment
of larger subunit of
ribosomes 50s
prokaryotes
in

60s
eukaryotes.
and in

2. CHAIN
ELONGATION:
ar
means
e

the
After of
formation complete
ribosome mRNA-tRNA

complex, an amino
alye
site (A-Site) establishes
acception
next to p-side (Peptide site).
 -
mRNA codon next
exposes
to initiation codon.

↓
A tRNA
aminoacye
new

reaches A
complexe site

and codon-anticodon
forms
bonding
↳

requires elongation
This
bonding
factor energy GTP.
and i.e.

A
formed
peptide
bond is

cool
between
group of first
acid
amino (Methionine) and

Nh second amino
group of
acid.

changed tRNAs
two are
If
->

enough, the formation of

close

peptide bond would be favoured

 ↳
enhances
Presence
of catalyst
-

the
of peptide
formation
bond
formation.

> Enzyme- PEPTIDYL

TRANSFERASE

(type of ecibozyme)

Here,
elongation factors
are

required.

TRANSLOCATION movement
of
-

ribosome on mRNA.

Ribosome move
from one codon

to the mRNA.
along
net

↓
Amino added
get
acids one

by one.

↓
Translated into
polypeptide
sequences
 dictated
by DNA, represented
RNA-
by
A translational
unit inmRNA

sequence of
RNA
flansed
-

the start and

by codon Aug
the
stop codon and codes
for a

polypeptide.
An mRNA also has some additional
that
donot translated
sequences get
↓
These are UTR's

(Untranslated
regions
A
at both 5'end and
percent
also at send after coden.
stop
↓
These are
for
required
EFFICIENT TRANSLATION

PROCESS,

3. CHAIN TERMINATION:

-
The termination
of polypeptide
 -
three
signalled by one
of the

termination codons UAA, UAG,

UGA.

A known
GTP-dependent factors
release associated
as
factor
is

termination
with codon.

In eRF,
eukaryotes
Prokaryotes RF, and RFz.

↓
translation
help terminating
in

the
and
releasing complete
polypep-
tide the
from ribosome.
-
REGULATION OF
GENE EXPRESSION:

Regulation
of gene expression
-
broad

term occurs at levels.

various

In levels:
eukaryotes, four
1.
Transcriptional
level -

of
Formation

transcript.
2.
Processing
level -

Regulation of splicing.
3.
Transport
of mRNA
from nucleus to the

cytoplasm.
4. Translational level.

The to
genes the
in
all-expensed
perform a
particular function
or

set
a
of functions.

In
eukaryotes
related
functionally genes
donot but
represent an
operon
a re

present
at different
sites.

↳
Development
and
differentiation
of
adult
into
embargo organisms
are also

a result
of
the coordinated
regulation
of
of
expression several sets
of genes.

->
It
metabolic, or
physiological
is

environmental condition
that
regulates
the the
expression of gene.
Genes ConstueveReNE (easier
 OPERON CONCEPT

nee
Francois Jacob Monod
and
Jacques
I (a biochemist)
geneticist)
a

proposed a model
of gene regulation.
↓

called model bacteria.

operon
an in

Operon-co-ordinated
group of genes
such as structival
gene,
operator
gene, promote
gene,
regulator gene functions
which

together and
regulate
an

pathway
metabolic as a unit.

↓
operon, top operon,
Ex: lac

his
are
operon, operon
val
operon.

-genes:
:Regulator gine: synthesise a biochemical
 which
or
protein
regulator
as
can act
positively
negatively
activation and as

expression.
:It the
controls
activity
of operation
gene.

2.
Operator gene:Gene the
receives
which

product
of regulator gene.
functioning
Se allows the

of the
openon
when not

covered biochemical
by the

produced by regulator gene.

3. Promote attachment
Provides
gene:
RNA
for polymerase.

n. Structural Transcribes mRNA

gene.
for polypeptide
synthesis.

THE LAC OPERON:

nee

lac-lactose
 Lac
operon:
I
gene/inhibitor
regulatory gene
(i)

I
promotergene
I
operation gene
3 structural A
gener polycistnoic
-

gene by
regulated
a common

and
promotion regulatory
gene.

In breakdown lactose
E.coli, of
requires enzymes.
three

↳
a
together
synthesised in co-ordinated

unit
manner
by functional of
DNA lac
operon.

The addition
of lactosestimulates
itself
the
production of enzymes
required
... it
iscalled inducible

system.

-on
genes:
1. STRUCTURAL GENES:
nee
 ->
lac
z:
codes
for B-galactosidase
-

the
is
which
responsible for
hydrolysisof disaccharide
the

i.e -

lactose-Galactose +

gencose
-

unit.
monomeric

lacy:-codes for permease.

->

increases
permeability
-

of cell B-galactosi-
the to

des

->
lac a:-codes
for transactylase
can
which
transfe
group -galactoside
acetyl
to

2. OPERATOR GENE:
-en

molecule
-

interacts
with
a
protein
or
regulator molecule which
prevents
 structional
the transcription
of genes.
3. PROMOTER GENE:

-gene possesses site

for
RNA
polyme-
attachment.
-

rase

4. REGULATOR (i)
gene
codes
gene for protein known
-

as Represson protein
↓
it
is
synthesized all the from
time

why
in it is
gene-that
is

which
constitutive
gene is
always
functional.

OPERON -
OFF:
SWITCHED

when
represson protein produced by
to
binds
regulatory
or inhibitor
gene
operator
gene
RNA
polymerase blocked.
. -

Thus, no
transcription.

Represson protein Operation gene

+

I
 ↓
switered
off

Regulation
of lac
open by now
-
control
negative
or
regulation.

If lactess-provided in the
growth
medium
of lactose
bacteria is

transported the
through of
action

permease.

low
->

Avery level
of
of expression
las that to be
present
in
operon
the cell all the time.
otherwise,
lactose cannot the
enter cell.

inducer laction
of
In such as
presence
repressor
the
or allolactose is

inactivated interaction
with
by
inducer This
allows RNA

to
polymerase access
promote and

transcription
proceeds.
 genome project (HGP):
thuman

Hap was the collaboratives

international
/

goal
research whose the
program was

compete
mapping
and
understanding
of
all
genes of human
beings.

HGp- "mega project" 13

year project.

co-ordinated US
by the
Department
and Institute
of Energy National
the

Health.
of
 Then Welcome Trust (UK) joined
the
project as
major partner,
additional
contributions
from
Japan, France,
Germany,
China

and others.

Project was
completed 2003.
in

Way +Gp
mega project?
estimated 9 US
billion
1.
Huge cost

dollars.

(cost US$3.
of sequencing (bp ->

Very large pains

2. number
of base

(3x109bp) to be
identified
and
-

sequenced.

3.
Requires a
large number
of
scientists, technicians and

supporting
staff.

4.
Storage of data:

some 3300 books.

1000
appeare pages each.
 typed
-
each has 1000
page
letters

High-speed computational
devices
for
retrieval
storage, and
analysis
of
data made it to
easier do the same.

5. Science
of
also
Bioinformatics developed
this
during period
and
helped 16p.

-HGP:

Identification
of all
approximately
-

20,000 25000 human DNA.

genes
-

in

Determine
the 3 billion
of
the
sequences
-

chemical base that

make
pains up
human DNA.

information
- To this
store in databases.

-
To
improve tools
for data analysis.
Transfer-related
technologies
to other
-

sectors, such as industries.

51: legal and

To
-
solve ethical,
any
issues.
social

association
-

Bioinformatics close

development of
a

of HGP with rapid

new area
biology.
in
-me again in
model ene

can lead to an
of
understanding
natural
their that
capabilities
be towards
applied solving
can

health-care,
challenges in
agriculture,
energy production,
environmental

remediation.
·
Non-human model such
organisms
as bacteria,
yeast,
Caenorhabditis
elegans
I a
non-pathogenic rematodel,
free-living
Deosophila, plants
like
rice and

etc.
Arabidopsis have been
sequenced.

dologies: Two
major approach:

ESTs
Expressed
1.
sequence Tags:
-

Identifying
all that
a re

genes
expressed as RNAs.

the whole
Sequence Annotation:
sequencing
2.

n e e

contained
that
set
of genome
all the and
coding non-coding
 assigning
and later
sequences

regions
different sequence
in

functions.
with

process:
sequencing
Total
DNA -isolated

cell
from
a

↓
converted into random

fragments of relatively
smaller
sizes.
↓
using
closed suitable

vectors.
specialised
↓
into
coming
results

each
of
amplification

piece of DNA
fragment
so that
it
subsequently
could be
sequenced
with
ease.

Bacteria
used bests
commonly and
yeast.
 vectors BAC

-
Bacterial artificial
chromosome.

X AC

to
-east
artificial
chromosome.

Fragments sequenced using

automated DNA
sequencers
that worked on the
perinciple
developed
method
by
Frederick

also
sanger, for
credited

method
developing
determi
for
-nation
of acid
amino
sequence

in
proteins.
These
->

were based on
sequences
some
regions
overlapping present
in them. For this,
of
generation
overlapping
fragments were

for sequencing.
required

↓
this
since isnot
possible
humans,
in specialised computer
 were
based
programs
developed

sequence of chromosome I
completed
was

only May
in 2006.

22 autosomes and X andy -

to

be
sequenced.

Features
of human
Genome:
->

contains
3164.7 nucleotide
million

bases.

Average gene
->
consists 3000 bases.
of
Largest
brown
size varies.
gene
DYSTROPHIN-2.4 million
bases and

TDF smallest with

14
gene
bases.

->
Total number
estimated
of genes
at
30,000- much lower than

estimates 80,000 to
1,40,000
precious
Almost 99.9% nucleotide bases
genes.
are
exactly
same in all
people.

over
-> are
Functions unknown for
50% discovered
genes.
of
-> Less than I
percent
of generesfor
proteins.
->
Repeated sequences make
up very
human
large portion of the genome.
Repetitive
sequences are stretches
->

of DNA
sequences
that are
repeated
times some 100 1000
to times.
many
-

They have no direct

function
coding
but chromosome
shed
light
on

and evolution.
structure, dynamics
->
chromosome 1 most (2960)
gener
I the
fewest-251.
->
has

->
Scientists
identified
have about

1.4 million
locations
where
single
pair
base DNA
differences
occur

humans.
in

↳ These are SNPs

Single
-

nucleotide
polymorphisms-snips
This
information
promises
revolution
to -

chromosom
-

the
is
process of finding
 al locations disease-associated
-

for
sequences
and human
tracing history.

ions and
-
CHALLENGES:

1
Completion
of first phase of human

been
genome project compared
has

antibiotics
because
discovery of
to

base
it has
opened a vast data
of
knowledge
about
various
aspects
of human
genome.
Soon shall be
mapping
2. we

all human
the all
genes,
junk
and
sequences, transposons
DNA.

3. More 1200
than that
cause
genes
common cardiovascular ailments
/

disease
endocrine diabetes,
like

Alyhemier's diseases, cancers, other

ailments.

After snapshot-will
taking to
be to know metered
the
possible
 alter them and remove
posibility.
4.
Simple produce
a

gene defects
-

that
of hereditary
number diseases

can be corrected.

interactions
blu
Possible
study
5. to

genes, proteins
various and the

mechanism
of forming tissues,
tumors
organs,
etc

Holds healthier and

6.
promise of
longer living.

Fingerprinting
DNA

Technique used
for nucleotide
determining DNA-
sequences of areas
certain
of
to each individual.
unique

fingerprinting-can distinguish
DNA

human
one
from
being
anothe

Exception Monozygotic
twins.
-

humans
99.9% base
of sequences among
 is the same.

3x106
They
have
of genere on

in
differences base
sequence.

fingerprinting
involves
identifying
DNA

differences
in some
specific
regions
in

DNA called REPETITIVE DNA.

sequence

These
repetitive DNA-
from
separated
bulk DNA
genomic different as
peaks
density
during gradientcentrifugation.
Bulk
DNA-major peak.
small
peaks-satellite
[
DNA.

-
I
into the
classified
categories
following
an

the basis
of:
!Base (A:T G:1)
composition or
rich

Length
of segment
2.

3. Number units.
of repetitive
CATEGORIES:
1. Rs-variable number
of
Tandem Repeats
or

MINI SATELLITES.

A
- arranged
small DNA
sequence
many
in
tanderly
numbers.
copy
copy from
number-varies chromo-

-some to chromosome in an

individual.

Number
of repeats show
very
*

high degree of polymorphism.

->
0.1 20kb
to

size of UNTR ->

(Kilobase)

SSRS
Single Sequence
2.
Repeats
-

STRS-Short
or
Tandem
Repeats
M
ICROSATELLITES

(1
6-p).
-

Basis
of fingerprinting
DNA - UNTR

Isnows

high degree of
polymorphisms)
 of printing
finger
DNA was
Technique

by Jeffreys.
the
given
DNA involves Southern
fingerprinting
blot and
hybridisation uses radio-

labelled VNTR
- as
probe.

Steps:
Isolation DNA
of
↓

Digestion
of DNA
using
endonucleases
restriction

of
Separation
DNA
fragments
by
ELECTROPHORESIS/ RFLP

(Restriction

i
Leugenie
Fragment

↓
Transfer of separated
DNA to
fragments synthetic
membranes (nitrocellulose
or
nylon)
to
 labelled
Hybridisation
using
UNTR
proble
↳
DNA
of hybridised
Detection

by autoradiography.
fragments
↓
gives
Auto bands
radiogram
of different sizes
↓
These bands a
give
characteristic
pattern for
an individual DNA.

senstivity been
by
->
has increased
of
use

PCR.

Applications:

Paternity
1.
maternity
disputes.
-

2. Criminal
identification
Personal
3.
identification
4.
immigrant.
Close relations
of intending