MOLECULAR BASIS OF

INHERITANCE
 DISCOVERY OF GENETIC
MATERIAL - TRANSFORMING
PRINCIPLE
• In 1928, Frederick Griffith, in
a series of experiments with
Streptococcus pneumoniae.
• Two strains – S & R strain
• When Streptococcus pneumoniae
(pneumococcus) bacteria are grown
on a culture plate, some produce
smooth shiny colonies (S) while
others produce rough colonies (R).
• This is because the S strain
bacteria have a mucous
(polysaccharide) coat & were
virulent, while R strain does not.
• Mice infected with the S strain
(virulent) die from pneumonia
infection but mice infected with
the R strain do not develop
pneumonia.
• Conclusion
• R strain bacteria had somehow been
transformed by the heat-killed S
strain bacteria. The heat-killed S
strain, had enabled the R strain to
synthesise a smooth polysaccharide
coat and become virulent. This is
‘transforming principle’.
 BIOCHEMICAL
CHARACTERISATION OF
TRANSFORMING PRINCIPLE
• Oswald Avery, Colin MacLeod
and Maclyn McCarty (1933-44).
• They chose protein-digesting
enzymes (proteases) and RNA-
digesting enzymes (RNases) &
DNA digesting enzymes
(DNases).
• Heat killed S strain were treated
with each &then cultured with non
virulent R strain & injected into
mice.
• Proteases –- Heat killed S strain -–
Cultured with non virulent R strain
–- Mice –- Dead
• RNases –- Heat killed S strain -–
Cultured with non virulent R strain
–- Mice –- Dead
• DNases –- Heat killed S strain -–
Cultured with non virulent R strain
–- Mice –- Alive
• CONCLUSION
• Proteases & Rnases didn’t
affect the transformation of
non virulent R to virulent one.
• Digestion with DNase did
inhibit transformation,
suggesting that the DNA
caused the transformation.
• They concluded that DNA is
the hereditary material, but
not all biologists were
convinced.
 THE GENETIC MATERIAL IS DNA –
HERSHEY – CHASE EXPT
• The proof that DNA is the genetic
material came from experiments of
Alfred Hershey and Martha Chase
(1952).
• They worked with viruses that infect
bacteria called bacteriophages.
• The bacteriophage attaches to the
bacteria and its genetic material then
enters the bacterial cell.
• Protein contain sulphur, & no phosphate
• DNA contain phosphate instead of
sulphur
• The bacterial cell treats the viral
genetic material as if it was its
own and subsequently
manufactures more virus
particles.
• Hershey and Chase worked to
discover whether it was protein
or DNA from the viruses that
entered the bacteria.
• They grew some viruses on a
medium that contained
radioactive phosphorus and some
others on medium that contained
radioactive sulfur.
• Viruses grown in the presence
of radioactive phosphorus
contained radioactive DNA but
not radioactive protein because
DNA contains phosphorus but
protein does not.
• Similarly, viruses grown on
radioactive sulfur contained
radioactive protein but not
radioactive DNA because DNA
does not contain sulfur.
• Radioactive phages were
allowed to attach to E. coli
bacteria.
• Then, as the infection
proceeded, the viral coats were
removed from the bacteria by
agitating them in a blender.
• The virus particles were
separated from the bacteria
by spinning them in a
centrifuge.
• CONCLUSION
• Bacteria which was infected with viruses
that had radioactive DNA were
radioactive, indicating that DNA was the
material that passed from the virus to
the bacteria.
• Bacteria that were infected with viruses
that had radioactive proteins were not
radioactive.
• This indicates that proteins did not
enter the bacteria from the viruses.
• DNA is therefore the genetic material
that is passed from virus to bacteria
 PROPERTIES OF GENETIC
MATERIAL
(i) It should be able to generate its
replica (Replication).
(ii) It should be stable chemically
and structurally.
(iii)It should provide the scope for
slow changes (mutation) that are
required for evolution.
(iv)It should be able to express
itself in the form of 'Mendelian
Characters’.
 THE DNA
• DNA – deoxyribonucleic acid – is
the genetic material – most
abundant one
• First discovered by Friedrich
Meischer (1869) & named as
nuclein
• DNA is a polynucleotide having 2
polynucleotide chains
• There are two types of nitrogenous
bases –
1. Purines - Adenine and Guanine
2. Pyrimidines - Cytosine, Uracil and
Thymine
• Nucleoside – Sugar + Nitrogen base
• A nucleotide has three components
– a nitrogenous base, a pentose
sugar (deoxyribose for DNA), and a
phosphate group.
• Polynucleotide – Many nucleotides
joined to one another by
phosphodiester bond
• CHARGAFF’S RULE
• Erwin Chargaff observed that for a
double stranded DNA, the ratios
between Adenine and Thymine and
Guanine and Cytosine are constant
and equals one.
• A+G = C+T – 1:1 ratio
Double stranded polynucleotide chain
• The salient features of the
Double-helix structure of DNA are
as follows:
• Proposed by James Watson &
Francis Crick in 1953
(i) It is made of two polynucleotide
chains, where the backbone is
constituted by sugar-phosphate,
and the bases project inside.
(ii) The two chains have anti-parallel
polarity. It means, if one chain
has the polarity 5‘-3', the other
has 3‘-5'.
(iii) The bases in two strands
are paired through hydrogen
bond (H-bonds) forming base
pairs (bp).
• Adenine forms two hydrogen
bonds with Thymine from
opposite strand and vice-versa.
• Guanine is bonded with
Cytosine with three H-bonds.
• As a result, always a purine
comes opposite to a pyrimidine.
(iv) The two chains are coiled in
a right-handed fashion.
• The distance between a bp in a
helix is approximately 0.34 nm.
• There are roughly 10 bp in each
turn.

• The pitch of the helix is 3.4 nm

(a nanometre is 10-9 m)
DNA double helix
• The length of DNA is usually defined
as number of nucleotides (or a pair of
nucleotide referred to as base pairs)
present in it.
• Haploid content of human DNA is 3.3 ×
10ꝰ bp.
• Bacteriophage known as φ ×174 has
5386 nucleotides
• Bacteriophage lambda has 48502 base
pairs (bp)
• Escherichia coli has 4.6 × 106 bp,
• If the length of E.coli DNA is
1.36mm, Calculate the number of
bp?
• Distance between two base pairs =
0.34 nm (0.34×10‾⁶ m)
• Length of DNA double helix = 1.36
mm
• No. of bp = 1.36mm/0.34x10‾⁶mm
= 4x10⁶ bp
(v) The plane of one base pair
stacks over the other in double
helix. This, in addition to H-
bonds, confers stability of the
helical structure
 PACKAGING OF DNA HELIX in
Prokaryotes
• In prokaryotes, they do not
have a defined nucleus, the
DNA is not scattered
throughout the cell.
• DNA (being negatively charged)
is held with some proteins (that
have positive charges) in a
region termed as ‘nucleoid’.
• The DNA in nucleoid is
organised in large loops held by
proteins.
 PACKAGING OF DNA HELIX in
Eukaryotes
• There is a set of positively charged
proteins called histones made of
amino acid residues of lysine &
arginine.
• Histones are organised to form a
unit of eight molecules called
histone octamer.
• The negatively charged DNA is
wrapped around the positively
charged histone octamer to form a
structure called nucleosome.
Nucleosome
• A typical nucleosome contains
200 bp of DNA helix.
• Nucleosomes constitute the
repeating unit of a structure in
nucleus called chromatin,
threadlike stained bodies seen
in nucleus.
• The nucleosomes in chromatin
are seen as ‘beads-on-string’
structure when viewed under
electron microscope
• In a typical nucleus, some
region of chromatin are loosely
packed (and stains light) and
are referred to as
Euchromatin.
• The chromatin that is more
densely packed and stains dark
are called as Heterochromatin.
• Euchromatin is said to be
transcriptionally active
chromatin, whereas
heterochromatin is inactive.
• The packaging of chromatin at
higher level requires additional
set of proteins that
collectively are referred to as
Non-histone Chromosomal
(NHC) proteins.

• DNA – Nucleosome –
Chromatin – Chromosome
• How many nucleosomes may
present in a mammalian cell?

• No. of bp in a DNA of a
mammalian cell = 6.6x10ꝰ
• No. of bp in a nucleosome = 200
bp
• Nucleosomes present in the DNA
of a mammalian cell =
6.6x10ꝰ/200
= 3.3x10⁷
 RNA WORLD
• RNA was the first genetic
material.
• RNA used to act as a genetic
material as well as a catalyst
• RNA being a catalyst was
reactive and hence unstable.
• Therefore, DNA has evolved
from RNA with chemical
modifications that make it more
stable.
• Have four nitrogen bases –
Adenine, Uracil, Guanine,
Cytosine
• RNA being unstable, mutate at
a faster rate.
• RNA act as a genetic material
in some viruses – Tobacco
Mosaic Virus, Rabies
 REPLICATION – DNA to DNA
formation
• Mechanism of DNA Replication
• Unwinding of a small part of DNA
double strand with the help of
Helicase enzyme
• DNA dependent DNA polymerase
attached on the 3’ end of parent or
template strand of DNA with the
help of RNA primer
• DNA polymerization (synthesis of
DNA) begins 3’ – 5’ direction of the
parent strand ( 5’ – 3’ direction in
newly synthesized strand.
• New strand with polarity 5’- 3’ is
synthesized from parent strand
with 3’ – 5’ polarity which is
continuous & known as continuous
strand (leading strand)
• Another new strand with polarity
5’-3’ is synthesized from parent
strand with 5’-3’ polarity which is in
the form of small fragments known
as discontinuous strand ( lagging
strand)
 Watson-Crick model for
semiconservative DNA replication
• Small fragments of DNA in the
discontinuous strand is termed as
Okazaki fragments
• Okazaki fragments are joined by
DNA ligase which make the strand
continuous
• After the completion of replication,
each DNA molecule would have one
parental and one newly synthesised
strand.
• This scheme was termed as
semiconservative DNA replication
replicating fork
REPLICATION FORK
• Replication occur within a small
opening of the DNA helix
ORIGIN OF REPLICATION (Ori)
• Specific point where replication
occurs
RNA Primer
• Short stretch of RNA
• Francis Crick proposed the
Central dogma in molecular
biology, which states that the
genetic information flows from
• DNA – RNA - Protein.
• DNA replication is termed as
semi conservative. Why?
• After the completion of
replication, each DNA molecule
has one parental & one newly
synthesized strand. This
scheme was termed as
semiconservative DNA
replication.
• Meselson and Stahl’s Experiment:
Experimental Proof
• Matthew Meselson and Franklin
Stahl performed the following
experiment in 1958:
(i) They grew E. coli in a medium
containing ¹⁵NH₄Cl (¹⁵N is the
heavy isotope of nitrogen) as the
only nitrogen source for many
generations.
• The result was that ¹⁵N was
incorporated into newly
synthesised DNA
• This heavy DNA molecule could be
distinguished from the normal
DNA by centrifugation in a cesium
chloride (CsCl) density gradient
(ii) Then they transferred the cells
into a medium with normal ¹⁴NH₄Cl
and took samples at various
definite time intervals as the cells
multiplied, and extracted the DNA
that remained as double-stranded
helices.
• The various samples were
separated independently on CsCl
gradients to measure the densities
of DNA
• Thus, the DNA that was extracted
from the culture one generation
after the transfer from ¹⁵N to
¹⁴N medium [that is after 20
minutes; E. coli divides in 20
minutes had a hybrid or
intermediate density.
• DNA extracted from the culture
after another generation [that is
after 40 minutes, II generation
was composed of equal amounts of
this hybrid DNA and of ‘light’
DNA.
• Taylor & his colleagues (1958)
done a similar experiment in faba
beans (Vicia faba) by using
radioactive thymidine to prove
DNA replication in chromosome is
semi conservative
 TRANSCRIPTION
• The process of copying genetic
information from one strand of
the DNA into RNA is termed as
transcription.
• Template strand – DNA strand
with polarity 3’ – 5’ which code for
RNA
• Coding strand – DNA strand with
polarity 5’ – 3’ which donot code
for RNA
• DNA DEPENDENT DNA
POLYMERASE
• Catalyze the formation of RNA
• It can only polymerase the RNA
in one direction i.e, 3’ – 5’
direction of the template strand
• There is a convention in defining
the two strands of the DNA in
the structural gene of a
transcription unit.
• Since the two strands have
opposite polarity and the DNA-
dependent RNA polymerase also
catalyse the polymerisation in
only one direction, that is, 5'→3',
the strand that has the polarity
3'→5' acts as a template, and is
also referred to as template
strand.
• The other strand which has the
polarity (5'→3') and the
sequence same as RNA (except
thymine at the place of uracil),
is displaced during
transcription.
• This strand (which does not
code for anything) is referred
to as coding strand.
• All the reference point while
defining a transcription unit is
made with coding strand.
• To explain the point, a hypothetical
sequence from a transcription unit is
represented below:

• 3'-ATGCATGCATGCATGCATGCATGC-5'
Template Strand
• 5'-TACGTACGTACGTACGTACGTACG-3'
Coding Strand
 TRANSCRIPTION UNIT
• A transcription unit in DNA is
defined primarily by the three
regions in the DNA:
(i) A Promoter
(ii) The Structural gene
(iii) A Terminator
• The promoter and terminator
flank the structural gene in a
transcription unit.
• Promoter is said to be located
towards 5' -end (upstream) of the
structural gene (the reference is
made with respect to the polarity
of coding strand).
• It is a DNA sequence that
provides binding site for RNA
polymerase, and it is the presence
of a promoter in a transcription
unit that also defines the
template and coding strands.
• The terminator is located
towards 3' -end (downstream)
of the coding strand and it
usually defines the end of the
process of transcription
• Located at 5’ end of template
strand
• Site where transcription
terminates
• Structural gene- Transcribing
(copy out) region of DNA
• Why both the strands are not
copied during transcription?
1. If both strands act as a
template, they would code for
RNA molecule with different
sequences which resulting in the
formation of different proteins.
This would complicate the genetic
information transfer.
2. The two RNA molecules if
produced simultaneously would
be complementary to each
other, hence would form a
double stranded RNA. This
would prevent RNA from being
translated into protein.
MECHANISM OF
TRANSCRIPTION
• INITIATION: RNA polymerase
binding on promoter region of the
template strand with the help of
an initiation factor called sigma
factor & thus transcription begins.
Once the process initiates, the
initiation factor dissociate
• ELONGATION: Following the rule
of complementarity, polymerization
(synthesis) of RNA strand occurs
by using nucleotide triphosphate as
substrate
• TERMINATION: When the RNA
polymerase reach the termination
site, a termination factor (rho
factor) attached to it which leads
to the dropping of RNA strand
from the RNA polymerase & the
process of transcription stops.
Process of Transcription in Bacteria
• NASCENT RNA: RNA formed
immediately through the process
of transcription
• In prokaryotes nascent RNA itself
acts as mRNA & donot need &
processing
• In eukaryotes nascent RNA is
termed as hnRNA (heterogenous
nuclear RNA) & need some
processing to become mRNA
• How hnRNA is converted into
functional mRNA?
• OR
• What is the additional complexity
of eukaryotic transcription?
• This is also termed as post
transcriptional modifications or
mRNA processing.
• RNA SPLICING
• hnRNA carries both introns &
exons
• The coding sequences or expressed
sequences are defined as exons.
• The exons are interrupted by
introns – unexpressed & non-
functional sequences.
• It should be removed by the
process called RNA splicing
• CAPPING
• Methyl guanosine triphosphate is
added to the 5’ end of hnRNA
• Tailing – Adenylate residue (200-
300 nucleotide) is added on the 3’
end of the hnRNA
Process of Transcription in Eukaryotes
TYPES OF RNA AND THE PROCESS
OF TRANSCRIPTION
• In bacteria, there are three major
types of RNAs:
1. mRNA (messenger RNA)
2. tRNA (transfer RNA)
3. rRNA (ribosomal RNA).
• All three RNAs are needed to
synthesise a protein in a cell.
• The mRNA provides the template, tRNA
brings aminoacids and reads the genetic
code
• rRNAs play structural and catalytic role
during translation.
• tRNA – carry amino acid from cytoplasm
to the ribosome during protein synthesis
• Cistron – segment of DNA which code
for a polypeptide
• Monocistronic – structural gene with one
cistron & seen in eukaryotes
• Polycistronic – structural gene with more
than one cistron & seen in prokaryotes
• In eukaryotes – classification &
function of RNA
(i) There are at least three RNA
polymerases in the nucleus
• The RNA polymerase I transcribes
rRNAs (28S, 18S, and 5.8S)
• The RNA polymerase II
transcribes precursor of mRNA,
the heterogeneous nuclear RNA
(hnRNA) – precursor of mRNA
• The RNA polymerase III is
responsible for transcription of
tRNA, 5srRNA, and snRNAs (small
nuclear RNAs).
PROKARYOTIC EUKARYOTIC
TRANSCRIPTION TRANSCRIPTION
Takes place in Takes place in
cytoplasm nucleus
Only one type of RNA Three type of RNA
polymerase is present polymerase is present
No mRNA processing mRNA processing
Polycistronic mRNA Monocistronic mRNA
No RNA splicing Need RNA splicing
Transcription & Transcription &
Translation coupled Translation is not
coupled
 GENETIC CODE
• It was George Gamow, a physicist,
who suggested that in order to
code for all the 20 amino acids, the
code should be made up of three
nucleotides (triplet code).
• This was a very bold proposition,
because a permutation combination
of 4ˆ3 (4 × 4 × 4) would generate
64 codons; generating many more
codons than required.
• The chemical method developed by
Har Gobind Khorana was
instrumental in synthesising RNA
molecules with defined
combinations of bases
(homopolymers and copolymers).
• Marshall Nirenberg - cell-free
system for protein synthesis
• Severo Ochoa – enzymatic
synthesis of RNA (polynucleotide
phosphorylase)
• Made a RNA molecule without
template DNA
• Developed checker board for
genetic code
The Codons for Various Amino Acids
• The salient features of genetic code
are as follows:
(i) The codon is triplet. 61 codons code
for amino acids and 3 codons do not
code for any amino acids, hence
they function as stop codons
(ii) Some amino acids are coded by more
than one codon, hence the code is
degenerate.
(iii) The codon is read in mRNA in a
contiguous fashion. There are no
punctuations.
(iv) The code is nearly universal: for
example, from bacteria to human
UUU would code for Phenylalanine
(phe).
(v) AUG has dual functions. It codes
for Methionine (met) , and it also
act as initiator codon.
(vi) UAA, UAG, UGA are stop
terminator codons
(vii) One codon code for only one
amino acid. Hence it is unambiguous
& safe
 tRNA– THE ADAPTER MOLECULE
• tRNA can code on one hand & other
hand would bind to amino acid
hence it is called an adapter
molecule
• tRNA has an anticodon loop that
has bases complementary to the
code, and it also has an amino acid
acceptor end to which it binds to
amino acids.
• tRNAs are specific for each amino
acid
tRNA - the adapter molecule
• For initiation, there is another
specific tRNA that is referred to
as initiator tRNA – provide first
amino acid for the process of
protein synthesis.
• There are no tRNAs for stop
codons.
• The secondary structure of tRNA
has been depicted that looks like
a clover-leaf.
• In actual structure, the tRNA is
a compact molecule which looks
like inverted L.
TRANSLATION
• The process of polymerisation of
amino acids to form a polypeptide
• Synthesis of protein from mRNA
• Takes place in ribosome
• Ribosome (prokaryotets) has two
sub units – larger (50S) & smaller
(30S)
• Amino acids are activated in the
presence of ATP and linked to
their cognate tRNA – a process
commonly called as charging of
tRNA or aminoacylation of tRNA.
• A translational unit in mRNA is
the sequence of RNA that is
flanked by the start codon (AUG)
and the stop codon and codes for
a polypeptide.
• An mRNA also has some
additional sequences that are not
translated and are referred as
untranslated regions (UTR).
• The UTRs are present at both 5'
-end (before start codon) and at
3' -end (after stop codon). They
are required for efficient
translation process.
• MECHANISM OF
TRANSLATION
i. Charging of tRNA
ii. mRNA – ribosome complex is
formed
iii. For initiation, the ribosome
binds to the mRNA at the start
codon (AUG)
iv. The ribosome proceeds to the
elongation phase of protein
synthesis.
v. During this stage, complexes
composed of an amino acid linked
to tRNA, sequentially bind to the
appropriate codon in mRNA by
forming complementary base pairs
with the tRNA anticodon.
vi. Amino acids are added one by
one, translated into Polypeptide
sequences by forming peptide bond
in between the amino acids
vii. At the end, a release factor
binds to the stop codon,
terminating translation and
releasing the complete
polypeptide from the ribosome.
• RIBOZYME
• Ribozyme act as catalyst. 23S
rRNA in bacteria is the enzyme
ribozyme
 CENTRAL DOGMA
Unidirectional flow of information
from DNA to mRNA and to protein
REGULATION OF GENE EXPRESSION
• Gene expression results only in the
formation of a polypeptide, it can be
regulated at several levels.
• In eukaryotes, the regulation could be
exerted at
(i) transcriptional level (formation of
primary transcript)
(ii) processing level (regulation of
splicing)
(iii) transport of mRNA from nucleus to
the cytoplasm
(iv) translational level
• OPERON CONCEPT – Used to
explain gene expression (Francoise
Jacob & Jacques Monod)
• OPERON – Group of structural gene
which is under the control of a
single gene promoter
• OPERON consist of –
• Structural gene – Portion of DNA
which transcribes into mRNA
• Promoter gene (P) – Site where RNA
polymerase binds to initiate
transcription
• Operator gene (O) – Region of DNA
which controls the structural gene
• Regulator gene (i) – Code for a
repressor protein
• Repressor – Protein coded by
regulatory gene which binds on
the operator & inhibit the binding
of RNA polymerase on it so that
the mRNA formation can be
controlled
• Inducer – A molecule that can
bind to the repressor so that the
RNA polymerase can be attached
on the operator & can initiate
mRNA synthesis
 THE LAC OPERON
• The elucidation of the lac operon
was by Francois Jacob and a
biochemist, Jacque Monod.
• In lac operon (here lac refers to
lactose), a polycistronic
structural gene is regulated by a
common promoter and regulatory
genes.
• The lac operon consists of one
regulatory gene (the i gene) and
three structural genes (z, y, and
a).
• The i gene codes for the repressor
of the lac operon
• The z gene codes for beta-
galactosidase (β-gal), which is
primarily responsible for the
hydrolysis of lactose into galactose
and glucose.
• The y gene codes for permease,
which increases permeability of
the cell to β-galactosides.
• The a gene encodes a
transacetylase – catalyse
transacetylation of lactose into
active form
• All the three gene products in lac
operon are required for
metabolism of lactose.
• Lactose is the substrate for the
enzyme beta-galactosidase and it
regulates switching on and off of
the operon. Hence, it is termed as
inducer.
The lac Operon
• In the absence of lactose (inducer)
regulator gene produces a repressor
protein binds to the operator region
of the operon and prevents RNA
polymerase from transcribing the
operon – switching off.
• In the presence of an inducer, lactose
bind to the repressor protein so that
the RNA polymerase can attach on
the operator & synthesize mRNA –
switching on
• This leads to the translation of mRNA
& production of B- galactosidase,
permease & transacetylase
• Regulation of lac operon by
repressor is referred to as
negative regulation.
• Lac operon is under control of
positive regulation.
MUTATIONS AND GENETIC CODE
• Sudden heritable changes in the
genotype of an organism

• POINT MUTATION
• Arise due to the change in the
single base pair of DNA
• Eg: sickle cell anemia.
• FRAMESHIFT INSERTION OR
DELETION MUTATIONS
• Insertion or deletion of one or two
bases changes the reading frame
from the point of insertion or
deletion.
• Insertion or deletion of three or
its multiple bases insert or delete
in one or multiple codon hence one
or multiple amino acids, and reading
frame remains unaltered.
• HUMAN GENOME PROJECT
• Launched in the year 1990.
• Completed in 2003
• Coordinated by US Dept. of
Energy & National Institute of
health
• Investment – 9 billion US$
 Goals of HGP
(i) Identify all the approximately
20,000-25,000 genes in human
DNA
(ii) Determine the sequences of
the 3 billion chemical base pairs
that make up human DNA
(iiii) Store this information in
databases
(iv) Improve tools for data analysis
(v) Transfer related technologies
to other sectors, such as
industries;
(vi) Address the ethical, legal, and
social issues (ELSI) that may arise
from the project.
• METHODOLOGIES
• The methods involved two major
approaches:
1. Expressed Sequence Tags:
Identifying all the genes that are
expressed as RNA (referred to as
Expressed Sequence Tags (ESTs).
2. Sequence Annotation:
Approach of simply sequencing the
whole set of genome that
contained all the coding and non-
coding sequence, and later
assigning different regions in the
sequence with functions
• For sequencing, the total DNA
from a cell is isolated and
converted into random fragments
of relatively smaller sizes
• Cloned in suitable host using
specialised vectors
• The commonly used hosts were
bacteria and yeast, and the
vectors were called as BAC
(bacterial artificial
chromosomes), and YAC (yeast
artificial chromosomes).
• The fragments were sequenced
using automated DNA sequencers
that worked on the principle of a
method developed by Frederick
Sanger.
• For alignment of these sequences
specialised computer based
programs were developed
• The sequence of chromosome 1
was completed only in May 2006.
• Genetic & physical maps were
generated using information on
polymorphism of restriction
endonuclease recognition sites,
and some repetitive DNA
sequences known as
microsatellites
A representative diagram of
human genome project
SALIENT FEATURES OF HUMAN
GENOME
• Some of the salient observations
drawn from human genome
project are as follows:
(i) The human genome contains
3164.7 million bp.
(ii) The average gene consists of
3000 bases, but sizes vary
greatly, with the largest known
human gene being dystrophin at
2.4 million bases.
(iii)The total number of genes is
estimated at 30,000–much
lower than previous estimates
of 80,000 to 1,40,000 genes.
Almost all (99.9 per cent)
nucleotide bases are exactly
the same in all people.
(iv) The functions are unknown for
over 50 per cent of the
discovered genes.
(v) Less than 2 per cent of the
genome codes for proteins.
(vi) Repeated sequences make up
very large portion of the human
genome.
(vii) Repetitive sequences are
stretches of DNA sequences that
are repeated many times,
sometimes hundred to thousand
times. They are thought to have no
direct coding functions, but they
shed light on chromosome
structure, dynamics and evolution.
(viii) Chromosome 1 has most genes
(2968), and the Y has the fewest
(231).
(ix) Scientists have identified
about 1.4 million locations where
single - base DNA differences
(SNPs – single nucleotide
polymorphism, pronounced as
‘snips’) occur in humans. This
information promises to
revolutionise the processes of
finding chromosomal locations for
disease-associated sequences and
tracing human history.
• Dystrophin – Longest DNA of
human body seen in X
chromosome having 2.4 million bp
• TDF – Testis Determining factor
– Smallest gene of humans seen in
Y chromosome having 14 bp
• VNTR – Variable number tandem
repeats or minisatelite- Smallest
DNA sequences arranged in a
DNA at specific intervals, it’s a
kind of satellite DNA having 11-
60 bp
• Repetitive DNA – DNA segment
having repeated sequence & it can
be transcribed
• Satellite DNA – Highly short
sequence in the repetitive DNA &
it cannot be transcribed
• Microsatelite (SSR – Simple
sequence repeats) – Another
form of satellite DNA having 5-8
bp length
• Polymorphism – Occurrence of
more than form of genetic
material
 DNA FINGERPRINTING
• Alec Jeffrey
• Probe- Radioactive labelled single
stranded DNA
(i) isolation of DNA
(ii) digestion of DNA by restriction
endonucleases
(iii) separation of DNA fragments by
electrophoresis
(iv) transferring (blotting) of
separated DNA fragments to
synthetic membranes, such as
nitrocellulose or nylon
(v) hybridisation using labelled VNTR
probe
(vi)detection of hybridised DNA
fragments by autoradiography.
Schematic representation of
DNA fingerprinting
• APPLICATIONS
• Used to settle parental dispute
• Used in forensic work
• Study evolutionary aspect