ACTATTCGATCCATAGCAT bil ion)

Human Genome Project

DNA
Fingerprinting
DNA C Nucleotide wiSedines
 Human Genome
Project/ Mega Project
known .
Mega Project
·

as
-
launched in 1990 (13 d-End-2003)
years
·

~
·

Estimated Genome Size 3

num
x109
Ap (3 billion)
- us
each $Ja
Cost
Estimated
·

3$ I -pl
Data 1000
19 billion
vi
Letters in each
page
·

Page ooe
-

1000 in each b

required200book
-

&
Data
~
devices
alysis required min speed computational
Emerge biologY
·
new a in i :
e

bioinformatics Application of usof computational

tools
-

analysis to
cof interpretation
I
of biological Data
-
.

US.
Department of Energy
· .

I of Health (VI)
National ·

Institute

Agencies
Involved .
-

· Welcome Trust (U.K)

Japan ~

countries
+
-
German

China-
~

France ~

Goals
of
~
HGP
-
-
Identify all the
approximately L
20000-25000 genes
·

in human DNA .

Chemical
Determine the
&
sequences billion I
·

ep human :
V
 ~

-
Store
information
this database I
·
.
-

Transfer
related
Technologies to other Sectors
&

&

industries
-

such
-

as
-
ethical
[
·
Address that
-
, a
legal
-
social
the
-
issues
(ELSI)
u
that
may
arise
from project -

unicellular
-
Bacteria
J i)

I
V

.
V Important Yeast v
ii)
Caenorhabditis elegans ~
i Jestwind
NonumanProsophiaI sea&
(Non-Parasitic ---- -Nematodes
-
S
in)

&

ArabidoIsis (Thale Crus plant)

NIv]
Methodologies
~

used
Ex Sequence Tags
-
------ - -
Sequence annotation

lESTs)
all S whole
Identify the
·

ages equence
·

that
-
expressed as RNE Genome
-

>
-
Expressed squence
Exon
-
O
Intran
+
-
-

mRNA
Steps 1) :
Total DNA
from a cell is .-
isolated
fragment. (DNAce/R-Ef
---

cleared
- -

ii) into small

ii) cloned into Bacteria/yeast
num
BAC S
using -

*C (Bacterial Artificial chromosomes S

·

Yeast
Artifical Chromosome) Vector
z
iv) Fragments Sequenced usingbanger Sequencer

by
--

-) Arranged in a
sequence
developed
based
delion

overlapping ~
sequence ↑

vi) specialised compwaterthesebased programs were

align
- -

developed to
sequences
A

Feature
Salient
of Human Genome

i) 3164 ·
7 million
bY
⑮Ap
&
- -

ii) Average gene

=
3000
bp ,
sige varies

Dystrophin (2 4 He
-

Largest gene
greatly
-

> .

iii) Total of genes

no 30 (Estimated 140000
=
, 000
-
80000 -

150 & % of D NA gene

total
u
functionNon is unknown
(
iv
X
~ (loding seam
-

v) Repeative
but
Present
coding
shed light on chromosome fre
·
-

function
-

Evolution
S

dynamic
Y

& -
.
-

vil Most Gene Chromosome 1 =

2968
genes
-

-
nu -
Least Gene = Chromosome Y =
231
nu
-
 Repetitive Yali-ship
:

vii) 2
% genome
code
for protein
114 million
-
-

viii) scientists identified #Ps T

ie

known Single Nucleotide

Polymorphism
Also
Snips
as

chromosomal
May help in
finding locations
·

for disease associated &

-
-
sequences
human
tracing
- history
-

-
~

Challenges :
~ -
~

i) Ass the J
genetic
I -
&
/physical maps
the igning
or

using Reinformation
-

on
genome sites
polymorphism of ~ recognition
↑

-
-

ii) Also microsatellites)

- are .
used
(Part Repetitive DNA)
Applications :

i) Deriving
Meaningful
g knowledge .
~
ii)
Understanding of biological systemsto
-
↑

iii) Enabling
a

radically
Research
approact new

biological -
~
iv) study interconnected genesin of geom me
by
Can a

Y can S

genes
- proteins S &
 >
-
How DNA appears
DNA
Fingerprinting etitive
-
DNA
↓
DNA)
known LRelATCATAT
Also
profiling is same
-
·
as DNA ChAT .

%99 9 DNA base

sequence among
.
↑

humans 10 01 %
unique)
-

· It

individuals identify -

genetic differences blu

two
&

DNA
fingertprinting identify ISpecific
DNA
·

known 3
sequence Repetitive
-
DNA

strect
* Refective DNA-- Small - repeats many
times
Form small peaks
fo
·

BuR DNA rm
major peak in
centrifugation
-

uks
(a)
2 lootings
ATGGETCGATCS -

(CTACTACTAA-1000 time (leght

--
W
S
mor 6 Sm

e Satellite DNA
Non, codinform
gno prote
-

Atype of
Defective Peaks
DNA
in
centrifugation
as

&
- - - -

Different basis
types on the
of
I
Composition of Dase
-

i) length of
T

segment
~

iii) Repeats
=

No
of ⑮
.

Different types -

t Misatellite Don't Code

for protea

ie
.
wigemm
Satellie
.
Variation at
his
due
-

to mutation .

V
 -
- -- D

Polymor
hisms are inheritable from parents
·

3
A

to childern
&

Polymorphism in
-
DNA -
Inheritable mutation
eopulation
a at
high
*
frequency .

role in
-

DNA
Polymorphism play important
·

evolution & peciation

S :
Radioactive substance
#
Toligonucleotide
#

DNA
Fingerprinting
= =
2 -
↑ -
4 8 -

w ↑

S- D
·

Develood b
Jeffreys (10-sep 6
Alec
1984)
which hous
,
-

Prepared - probe
sal Ye DNA - S
-

N- R
-
high degree of polymorphism UNT
very variable Rs F

repeats
O
&
ie
. number
offandom ation .

W7
nu
Earlier
-
he
-
used
radiolabelled
southern
-
-
-

UNTRs
hybridis
using
as a
probe
No is more- UNTR
VNTRs
·
-

Typof ini
-

satellite
:
aranged
DNA is
sequence in
copy number
: many
·
Show
-
high degree of polymorphism .

bige varies 0 1 to
from . 20kb

* 1) kb = 1000 nucleotide

Steps :

i) Isolation of D NA -
-
-
i) Endonuclease
Digestion of of GNA by DNA friction
iii)
- separation
electro ~ horesis
resegments by
fo
inf Transferring (
blotting) of separated -
DNA
fragment
 Probe >
-

Radio-active substance
=>
↓

oligonuleotide
! (short) 4-8 in no

-
-
#) MiaseTissue - ·
do -

↓ Digesta
↓

fat/lipid-breakdown is

enzyme
Cathode
Protease
- (seaweed
m
-

Agarose
-ve
- W
u

getGracelaria
u

↑ Wells ↑ -
e DNA-fragment
lovely-charged)
~ >

I7
-

>
O
-

D
3 A Ma a

-
~
- ~ VV
.
radiation
-

O
~
-
X T S

# Are Anode
Marker
--11

&
 to
syn
thetic membranes suc as
nitrocellulose)
or
-nylon
v Labelled >
Hybridisation using
UNTR robe .

of hybridised fragments
-

vif Detection DNA

"
autoradiography &

#
-

& DNA
-
same
fingerprint IMonozygotic
of wins -
are

Application :

Forensic Science v
&

Determing population & Genetic Diversities

-
V
33 X
:

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-

haploid (11)
3
:

3x109X2 >
-
6 6x109
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