RT

tes
6.0 Introduction
inheritance was not
At the time of Mendel, the nature of those 'factors' regulating the pattern of
nucleic acids
clear. Deoxyribonucleicacid (DNA)and ribonucieic acid (RNA) are the two types of
of the
(polymers of nucleotides) found in living systems. DNA acts as the genetic material in most alsa
organisms. RNAacts as a genetic material in some viruses, mostly tunctions as a messenger. RNA
functions as adapter, structural and in some cases as a catalytic molecule.
6.1 The DNA
DNA is a long polymer of deoxyribonucleotides. DNA is the macromolecule which consists of two
complementary strands of deoxyribonucleotides that run antiparallely and are held together by
hydrogen bonds between their opposite nitrogen bases. It consists of four types of basicunits called
nucleotides.The length of DNA is usually defined as number of nucleotides (or a pair of nucleotide
referred to as base pairs) present in it and is the characteristicof an organism.
For example, a bacteriophage known as Qx174 has 5386 nucleotides. Bacteriophage lambda has
48502 base pairs (bp), Escherichia coli has 4.6 x 106 bp and haploid content of human DNA is
3.3 × 109 bp.
Structure of Polynucleotide Chain
Anucleotide has three components - anitrogenous base, a pentose sugar (ribose in case ot RN
and deoxyribose for DNA) and aphosphate group. The nitrogenous bases are of two types - purines
(adenine and guanine) and pyrimidines (cytosine, uracil and thymine). Cytosine is common tor bom
DNA and RNA.Thymine is present in DNA and uracil is present in RNA at the place of thymine.
Anitrogenous base is linked to the OH of 1C pentose sugar through a N-glycosidic linkage to o
anucleoside. When a phosphate group is linked to 5-0H of a nucleoside throughphosphodiester
phosphodiegte
linkage, a corresponding nucleotide is forrmed. Two nucleotides are linkedthrough3-5"
linkage to form a dinucleotide. More nucleotides can be joined in such a manner to torti o
polynucleotide chain.
ribose. Also,
In RNA, every nucleotide residue has an additional -0H group present at 2 -position in the
in RNA the uracil is found at the place 5 phosphate H 3'hydroxyl
of thymine (5-methyl uracil, another P
H H

chemical name for thymine). DNA as an -oH

acidic substance present in nucleus was A
H
first identified by Friedrich Meischer G
in 1869 and named it as 'Nuclein'. Fig.: Apolynucleotide chain
Biology | Molecular Basis of Inheritance
173
Structure of DNA
In 1953, James Watson Francis Crick based on the y.rov
and
Rosalind Franklin, diffraction data nroduced by
was also based on theproposed a very simple but famous double helix model for the structure ofMaurice wikisad
DNA. The proposition
observation of Erwin Chargaff
which is
The purines and
pyrimidines are always in equal amounts, i.e.,summarised
A+
as :
that of thymine and the G=T+C.
The amount of adenine is always equal to
cvtosine, i.e., A = T and G = C. However, amount of quanine is always equai to lat o
The base ratio A + T/G+ C may vary
amount of A + Tis not
necessarily equal to g +
from one speries to another. but is
if each strand trom a DNA (parental DNA)
constant for a given species.
acts as atemplate for synthesis of a new
stranded DNA(daughter DNA) produced would be strand then tie two 0006
identical to the parental DNA molecule.
The salient features of the
() It is made of twO double-helix structure of DNA are as followS:
polynucleotide chains, where the backbone is constituted
by sugar-phosphate and the bases project inside.
(ii) The two chains have
anti-parallel
polarity 5 ’3',the other has 3polarity.
It means, if one chain has the Base pairs
’ 5'. Adenine Thymine
(i) The bases in twO strands are paired
forming base pairs (bp). Adenine formsthrough hydrogen bond (H-bonds)
two hydrogen bonds with thymine
from opposite strand and vice-versa. Similarly, Guanine Cytosine
quanine is bonded with
cytosine with three H-bonds. As a result, purine always
to a pyrimidine. This generates approximately uniform comes opposite Sugar phosphate
the two strands of the helix. distance between backbone

(iv) The two chains are coiled in a right-handed fashion. The pitch of
helix is 3.4 nm (a nanometre is one bllionth of a metre, that is 10-9 m)
the
Fig.: DNA double helix
and there are roughly 10 bp in each turn. Consequently, the distance between a bp in a helix is
equal to 0.34 nm. approximately
(V) The plane of one base pair stacks over the other in double helix. This, in addition to
the helical structure. H-bonds, confers stability of

H
P
H OH3
H

hydrogen
A bonds

OH
3

5
Fig.: Double stranded polynucleotide chain
Central dogma
Francis Crick proposed the central dogma in molecular biology, which states that the genetic information flows from
DNA->RNA ’ protein.
Replication
Transcription > mRNA ranslation
DNA >Protein
In some viruses, the flow of information is in reverse direction, that is, from RNA to DNA and is called
reverse central dogmalteminism or reverse transcription.
Transcription Translation
DNA RNA >Proteins
Reverse transcription
174 mtG Objective NCERT at your
Fingertips
Packaging of DNA Helix
>The length of DNA double helix in a typical mammalian cell is approximately 2.2 metres. Long sized DNA molerulae
are compacted in small areas only through packing.
DNA packaging in prokaryotes
> In prokaryotes, DNA is supercoiled (coiled and recoiled) with the help of RNAs and nonhistone basic proteins like
polyamines. The compacted mass of DNA is called nucleoid. NPP uchee m
DNA packaging in eukaryotes
DNA packaging in eukaryotes is carried out with the help of basic amino acids residues lysine and arginine called
histones.
Histones are organised to form a unit of eight molecules called histone DNA H, histone
octamer. The negatively charged DNA is wrapped around the positively
charged histone octamer to form nucleosome.
There are five types of histone proteins, i.e., H,, H,A, H,B, H, and H4. Four
of them (HzA, HzB, Hg and H¡) occur in pairs to produce histone octamer or Histone
nu-body. 0ctamer

Atypical nucleosome contains 200 bp of DNA helix. Nucleosomes constitute

the repeating unit of a structure in nucleus called chromatin (thread-like
stained bodies seen in nucleus). Core of histone molecules
The nucleosomes in chromatin appears as "beads-on-string' under electron Fig.: Nucleosome
microscope (EM). The beads-on-string structure in chromatin is packaged to form chromatin fibers that are further
coiled and condensed at metaphase stage of cell division to form chromosomes.
In atypicalnucleus, some region of chromatin are lo0sely packed (and stains light) and are referred to as euchromatin.
The chromatin that is more densely packed and stains dark are calied as heterochromatin. Euchromatin is said to
be transcriptionally active chromatin, whereas heterochromatin is inactive.
DNA connecting two adjacent nucleosomes iscalled interbead or linker DNA.It bears H,histone protein (called
plugging protein and act as marker protein). Nucleosome and linker DNA together constitute chromatosome.
 lac mRNA
Translation
Inducer B-galactosidase permease transcetylase
(lnactive repressor)
The repressor protein binds to the Fig.: The ac operon
operator region of the operon and prevents RNA polymerase from transcribing
the operOn. in tne presence of an inducer.
inducer. This allows RNA polymerase access(lactose or allolactocal the renressor is inactivated by
to the promoter and interaction with the
Requlation of lac operon by repressor is referred to as transcription procee0S.
negative regulation.
69 Human Genome Project
Human genome project (HGP) was launched in the vear
it is called international Human Genome 1990 and was called a mega project.
each of the human chromosomes determining Seguencing Consortium and is aimed at finding out all the genes in
their function and hopefully understanding how they together
complete organism. Ihe HGP was a 13-year project coordinated by the U.S. form the
Institute f Health. Human Department of Energy and the National
caled bioinformatics. Genome Project was closely associated with the rapid development of a new area in biology
The goals of the human genome project
are as follows :
1o deveiop a genetic linkage map of human genome by identifyingthousands of genetic
in the genome. markers and mapping them
To obtain aphysical map of human genome by cloning genomic DNA into YACs and
- To sequence the entire human
cosmids.
genome.
The two factors that made this possible are : () Genetic engineering techniques, which
and clone any segment of DNA and (ii) Availability of simple and fast techniques, for made it possible to isolate
determining the DNA sequences.
Methodologies of HGP
Expressed Sequence Tags (ESTs, 0.e., Focused on identifying all the genes that are expressed as RNA) and Sequence
Annotation. Sequence the whole set of genome, that included all the coding and non-coding sequences and later
aigring furnctions to different regions in the sequences are the methods used to sequence human genome
Salient Features of HGP
Some of the salíent observations drawn from human genome project are as follows:
6 The human genome contains 3164.7 million nucleotide bases pairs.
The ayerage gene consists of 3000bases, but sizes vary greatly, with the largest known human gene being
at 2.4 million bases. dystrophin
isy The total number of genes is estimatedat 30,000.Almost all (99,9 per cent) nuleotide bases are evacthb thsmo
ifn all people.
fM The functions of over 50 per cent of the discovered genes are unknown
 182
wtG Objective NCERT at
(v) Less than 2 per cent of the genome
your Fingertips,
codes for proteins.
(vi) Repeated sequences (stretches of DNA sequences that are repeated many times, sometimes hundred
to
times) make up very large portion of the human genome.
(Vi) Chromosome 1has most genes (2968) and the Yhas the fewest
(231).
(vii) Scientists have identified about 1.4 million locations where single base DNA differences (SNPs -
thousarnd
polymorphism, pronounced as 'snips') occur in humans. single nucleotide
Applications of HGP
Applications of HGP are as follows
(0) Having the complete
sequence of human genome, will
enable aradically new approach to biological
asystematicapproach on a much
broader scale.
(0) A he gene in a genome or all the transcripts in a
particular tissue/orqan/tumor can be studied.
research, i.e.
(ii) It will be possible to
understand
network in the chemistry of life. how the enormous number of genes and proteins work together in
6.10 DNA Fingerprinting
interconnected
DNA fingerprinting involves identifying
DNA, because in these sequences, a smalldifferences
in some specific regions in DNA
stretch of DNA is repeated many times. It is sequence
called as
called as repetitiye
tandem repeats (VNTR). The repetitive DNAS are separated from variable number of
gradient configuration. The bulk DNA forms amajor peak and the bulk genomic DNAas different peaks duringdensity
repetitive DNA sequence show high degree of other small peaks are called satellite DNA,
polymorphism and form the basis of DNA fingerprinting. The These
trom every tissue (such as blood, hair-follicle, skin, DNA
of poBymorphism, they become very useful bone, saliva, sperm etc.), from an individual show the
same degree
identification tool in forensic applications.
Polymorphism (variation at genetic level) arises due to mutations. In DNA polymorphism, more than one
(allele) at a locus oCcurs in human population with a
from extremely minute amounts of blood, frequency greater than 0.01. DNA fingerprints can be variant
semen, hair bulb or any other cells of the body. prepared
The technique of DNA fingerprinting was initially
shows very high degree of polymorphism. The technique, developed by Alec Jeffreys. He used a satellite DNA as probe that
radiolabelled VNTR as a probe. Major steps are as follows:as used earlier, involved Southern blot hybridisation using
DNA extracted from the
cells. DNA can be amplified by PCR or
DNA is cut into fragments with polymerase chain reaction.
restriction enzymes.
Chopped DNA fragments are passed through
them with a dye. electrophoresis. The separated fragments can be visualised by staining
Double-stranded DNA is then split into single-stranded DNA using alkaline chemicals.
Separated DNA sequences are transterred from gel onto a
(Southern blotting) HD nitrocellulose or nylon membrane
The nylon sheet is then immersed in a bath,where
probes or markers (radioactive synthetic DNA segments of
sequences) are added. The probe hybridises with VNTR. known
X-ray film is exposed to the nylon sheet gives dark bands at the
by autoradiography. probe sites. Thus, hybridised fragments are detected
DNA sequencing by Sanger's Method HD
DNAfingerprinting helps in identifying the true
(biological) father, criminal and it can solve the problems of evolution.