Class 12

Biology
Chapter 6: Molecular Basis of Inheritance

Apibeni B Bulsara

PGT-Bio

The DNA (Deoxyribonucleic acid)

• DNA is the genetic material in most of the organisms except some viruses, which have an RNA
genome, e.g. TMV (Tobacco mosaic virus)
• RNA mostly acts as a messenger, an adapter and has a catalytic function
• Number of base pairs (bp) or nucleotides determines the length of the DNA Human DNA
(haploid) – 3.3 x 109 bp
Bacteriophage 𝜙 x 174 – 5386 nucleotides
Bacteriophage 𝝀 – 48502 bp
E. coli – 4.6 x 106 bp Brief History

• Friedrich Meischer in 1869 identified DNA present in the nucleus as an acidic

substance and named it as ‘Nuclein’
• Frederick Griffith in 1928 demonstrated in the infected mice that some
“transforming principle” gets transferred from heat-killed Sstrain of
Streptococcus pneumoniae to R-strain, enabling it to make the smooth
polysaccharide coat, hence it becomes virulent
 Oswald Avery, Colin MacLeod and Maclyn McCarty determined the
biochemical nature of “transforming principle” and discovered that only DNA is
responsible for the transformation
• Hershey and Chase in 1952 proved that DNA is the genetic material. They
infected the bacteria E. coli with bacteriophages grown in radioactive phosphorus
(32P) which labels DNA of the bacteriophage, which gets transferred to the
bacteria cells and radioactive sulphur ( 35S), which labels the protein coat of the
bacteriophage, hence radioactivity is not detected in the bacterial cell.

Structure of a polynucleotide chain

• Each nucleotide is composed of three elements:
1. Nitrogenous base:
1. Purines- Adenine (A) and Guanine (G) present in DNA as well as
RNA
2. Pyrimidines- Cytosine (C) and Thymine (T) in DNA and Cytosine and
Uracil in RNA. Thymine is also known as 5methyl uracil and it is
accounted for more stability of DNA molecule
2. Sugar: Pentose sugar- Ribose in RNA (ribonucleic acid), deoxyribose in
DNA 3. Phosphate group
• Nucleoside: a nitrogenous base linked to the hydroxyl group of 1’ C of the
pentose sugar by N-glycosidic bond
Nucleotide: phosphate group is attached to the hydroxyl group present at 5’ C of
the nucleoside by a phospho-ester bond
 Structure of a Nucleotide

• 3’-5’ phosphodiester bond links two nucleotides together to form dinucleotide and
the chain continues to grow to form a polynucleotide

A Nucleotide Chain

Double Helix Model for Structure of DNA

• Watson and Crick in 1953 proposed the double helix structure of DNA
• Ervin Chargaff observed that the ratio of Adenine and Thymine to
Guanine and Cytosine is one and remains constant
• Two polynucleotide chains make DNA, where the backbone is of sugar-
phosphate and bases are present towards inside
• The two chains have the opposite polarity, i.e. one with 5’→3’ and the other has
3’→5’ polarity
Base pair (bp) is formed by hydrogen bonding between nitrogenous bases of
both the polypeptide chains
 • Always a purine base of one nucleotide chain is linked to a pyrimidine base of
another nucleotide chain or vice versa to make a base pair
• Adenine pairs with Thymine (or Uracil in RNA) by two hydrogen bonds (A=T)
• Guanine pairs with Cytosine by three hydrogen bonds (G)
• The two polypeptide chains are coiled in a right-handed direction
• The distance between two base pairs is 0.34 nm, there are 10 bp in each turn
and the pitch of the helix id 3.4 nm
• Stability of the helical structure is conferred by the presence of base pairs
stacked one above the other

The length of a DNA double helix is about 2.2 meters (6.6 x 109 bp x 0.34 x 10-
9
m/bp)
Therefore, it needs special packaging in a cell.
Packaging of Nucleotide:
• In prokaryotic cells (which do not have a defined nucleus), such as E.coli, DNA (being negatively
charged) is held with some proteins (that have positive charges) in a region called as nucleoid.
The DNA in nucleoid is organised in large loops held by proteins.
•In eukaryotes, there is a set of positively charged proteins called histones that are rich in basic
amino acid residues, lysines and arginines (both positive).
 • There are 5 types of histone proteins- H1, H2A, H2B, H3 and H4
• Histone octamer has 2 molecules of 4 histone proteins and it plays
an important role in gene regulation
• A nucleosome is a repeating unit in chromatin, it prevents DNA from
getting tangled
• Nucleosome contains around 200 bp of DNA
• Non-histone chromosomal proteins (NHC) help in further
packaging of chromatin
• Euchromatin: these are transcriptionally active areas, where
chromatin is loosely packed and they take up light stain
• Heterochromatin: these are transcriptionally inactive areas, where
chromatin is densely packed and they take up dark stain

Replication
• Watson and Crick suggested that the replication of DNA is
semiconservative
• Meselson and Stahl in 1958 experimentally proved that the
DNA replicates semi conservatively
(i) E. coli was grown in a medium containing 15NH4C1 as the only nitrogen
source for many generations. 15N got incorporated into newly synthesised
DNA (and other nitrogen containing compounds). This heavy DNA
molecule could be distinguished from the normal DNA by centrifugation in
a cesium chloride (CsCl) density gradient.

(ii) They then transferred the cells into a medium with normal 14NH 4Cl and
took samples at various definite intervals as the cells multiplied and
extracted the DNA that remained as double stranded helices. DNA
samples were separated independently on CsCl gradients to measure
DNA densities.
(iii) The DNA that was extracted from the culture, one generation (after 20
min) after the transfer from 15 N to 14N medium had a hybrid or
intermediate density. DNA extracted from the culture after another
generation (after 40 min) was composed of equal amounts of this hybrid
DNA and of light DNA.
(iv) Very similar experiments were carried out by Taylor and Colleagues on
Vicia faba (faba beans) using radioactive thymidine and the same results,
i.e. DNA replicates semiconservatively, were obtained as in earlier
experiments.
• Taylor et al in another experiment on fava beans (Vicia faba) using
radioactive thymidine proved that the replication on DNA is
semiconservative

Process of Replication.

• Enzyme DNA polymerase catalyses DNA replication. It can

polymerise only in 5’→3’ direction
• Replication is initiated at the origin of replication
• Deoxyribonucleoside triphosphate provides energy for the
polymerisation reaction and also acts as a substrate
• A small part of DNA opens up making a replication fork, where
replication occurs
• Replication is continuous in a strand with 5’→3’ direction, called
leading strand, where the template strand has 3’→5’ polarity, called
leading strand template
• Replication is discontinuous in the other strand, where the template
strand has 5’→3’ polarity, called lagging strand template
• The discontinuous fragments, called Okazaki fragments are joined
together by the enzyme DNA ligase
• In eukaryotic cells, the replication takes place during s-phase of the
cell cycle
• If cell division doesn’t occur after the replication, it results in
polyploidy of chromosomes
Transcription
• In the process of transcription, the genetic information present in the
DNA gets copied to RNA
• Only one segment of DNA gets copied to RNA
• In RNA Adenine pairs with Uracil instead of Thymine present in DNA
• Transcription of DNA includes three regions; a promoter, the
structural gene and a terminator
• RNA polymerase catalyses the transcription and direction of
transcription is same as that of replication by DNA polymerase, i.e.
5’→3’ direction
• Template Strand: it has 3’→5’ polarity, which acts as a template for
RNA formation, also called antisense strand
• Coding strand: it has 5’→3’ polarity, which has the sequence similar
to the newly formed RNA, except that Thymine is replaced by Uracil
in RNA, also called the sense strand
• Promoter: it is situated at the 5’ side of the structural gene or
upstream (with respect to coding strand). RNA polymerase binds
here to start the transcription
• Structural gene: the region between promoter and terminator.
Cistron is a segment of DNA that codes for a polypeptide. The
structural gene is monocistronic in eukaryotic cells and
polycistronic in prokaryotic cells
• Terminator: it is situated at the 3’ side of the coding strand and
marks the end of the transcription process
• The monocistronic structural gene (in eukaryotes) has interrupted
coding sequences, the gene in eukaryotes is called split genes
 • Exons: coding sequences, which appear in mature and processed
RNA
• Introns: intervening sequences. It is not present in the mature and
processed RNA
Transcription unit

Transcription in a Prokaryotic cell

• In bacteria, a single DNA-dependent RNA polymerase catalyses the
transcription of all the three RNAs present; mRNA, tRNA, rRNA
There are three steps of transcription:
1. Initiation: RNA polymerase binds to initiation factor, sigma
(𝞂) at the promoter site to start the process of transcription
2. Elongation: RNA polymerase is only capable of elongation
3. Termination: when the RNA polymerase reaches the
terminator region, it binds with the terminator factor, rho (𝜌) to
terminate the process and the nascent RNA falls off

Transcription process in Bacteria

 • In bacteria, mRNA doesn’t require further processing and as the
nucleus and cytosol are not separate, translation is coupled with
transcription and is initiated before the full transcription of mRNA
occurs

Transcription in a Eukaryotic cell

• There are three RNA polymerases to catalyse transcription of
different RNAs in a eukaryotic cell:
1. RNA polymerase I- transcription of rRNA (28S, 18S, 5.8S)
2. RNA polymerase II- transcription of hnRNA (heterogeneous
nuclear RNA), which is a precursor of mRNA
3. RNA polymerase III- transcription of tRNA, 5S rRNA and
snRNA (small nuclear RNA)
• The primary transcript is non-functional and has exons intervened by
introns
• Splicing- the process of removing introns and joining together exons
in a defined sequence
• Capping and tailing- additional processing of hnRNA. In capping a
methyl guanosine triphosphate is added to the 5’ end of hnRNA.
In tailing, 200-300 adenylate residues are added at 3’ end.
• mRNA is fully processed hnRNA, which gets transported out of the
nucleus for translation

Significance of these complexities are:

(i) The split gene arrangements represent an ancient feature of genome.
(ii) The presence of introns is reminescent of antiquity.
(iii) The process of splicing represents the dominance of RNA world.

Genetic Code
• It is the sequence of bases in mRNA, that codes for a particular
amino acid in the protein synthesis
• Each code is made up of three nucleotides called a triplet. codons
are nearly universal, except for some protozoans and mitochondrial
codons
• More than one triplet codon code for same amino acid, so the code is
degenerate
• There are a total of 64 codons, of which 61 code for amino acids
• 3 codons do not code for any amino acids, they are called stop
codons- UAA, UAG, UGA
• AUG is the start codon as well as codes for the amino acid
methionine

Ref to NCERT Text book for Genetic Code Genetic

Mutation
• Point Mutation: Change of single base pair results in the point
mutation, e.g. Sickle cell anaemia is a result of a point mutation in the
gene coding for the -globin chain. As a result Glutamate in the
normal protein gets converted to Valine in the sickle cell
• Frameshift Mutation: When there is loss or gain of one or two base
pairs, it changes the reading frame at the point of insertion or deletion
resulting in the frameshift mutation

Translation
• The translation is the process of amino acid polymerisation. Amino
acids are joined by peptide bonds
• All three RNAs have a different role in the process of translation
1. mRNA- provides the template. The sequence of amino acids in
a polypeptide chain is determined by the sequence of bases
present in mRNA
2. tRNA- acts as an adapter, it brings amino acids and reads the
genetic code
3. rRNA- performs a structural and catalytic role
tRNA – The Adapter Molecule:

Crick proposed the presence of an adapter molecule, which binds to a

specific amino acid. It was known as soluble RNA or sRNA and later
named as tRNA
• Shape of the tRNA is like an inverted ‘L’
• tRNA is specific for each amino acid, there is a specific initiator
tRNA
• There are no tRNAs for stop codons
•It has an anticodon loop, which has complimentary code present on
mRNA
• There is an amino acid acceptor arm, which binds the specific
amino acid as per the codon

• The first step in the process of translation is aminoacylation of

tRNA (charging of tRNA)
• Ribosomes are a protein manufacturing factory
• mRNA to protein translation begins with the presence of mRNA in the
small subunit of ribosomes
• The process of translation is always in 5’→3’ direction
• Peptide bond formation occurs between two amino acids present on
tRNAs in close vicinity
• There are two sites in the large subunit of a ribosome which
accommodates two tRNAs with amino acids close enough to form a
peptide bond
• Ribosome also acts as a catalyst in the formation of a peptide bond
• Start codon and stop codon flank the coding sequence for a
polypeptide in mRNA
• Untranslated regions (UTRs)- UTRs are present before the start
codon, i.e. at 5’ end and after the stop codon, i.e. towards 3’ end.
They are not translated but they make the translation process
efficient
• The Release factor binds to the stop codon at the end terminating
the process. The polypeptide gets released from the ribosome
 The Central Dogma
Francis Crick proposed that genetic information flows from DNA →
RNA → Protein, it is known as the central dogma of molecular
biology
Regulation of Gene Expression

• Expression of a gene to form polypeptide can be regulated at various

levels in eukaryotes
1. At the time of formation of a primary transcript, i.e. transcription
2. At the time of processing or splicing
3. At the time of transportation of mRNA from the nucleus to the
cytosol
4. At the time of protein synthesis, i.e. translation
• Environmental, physiological and metabolic conditions regulate the
gene expression
The development and differentiation of embryo is a result of
coordinated regulation and expression of several sets of genes In
prokaryotes, control of gene expression is mainly at the initiation of
transcription
• The activity of RNA polymerase at the start site is regulated by
regulatory proteins, which can be a repressor or activator
• The accessibility of the promoter region is regulated by an operator
sequence adjacent to it, that binds with the specific protein, mostly a
repressor
• In each operon, there is a specific operator and repressor protein

The Lac Operon

 • Jacob and Monad first showed the transcriptionally regulated system
in the lac operon
• An operon consists of many structural genes regulated by a common
promoter and regulatory gene
• The lac operon consists of
1. Regulatory gene: gene i (inhibitor gene), that codes for the
repressor of the lac operon
2. Structural gene: three structural genes, z, y, a
- z gene, codes for 𝜷-galactosidase, it hydrolyses
lactose to glucose and galactose
- y gene, codes for permease, responsible for increasing
permeability of the cell to 𝛽-galactosides
- a gene, codes for transacetylase
The repressor is synthesised continuously from the gene i, which
binds to the operator and prevents RNA polymerase from
transcribing
Lactose is a substrate of -galactosidase and also regulates the
gene expression. It acts as an inducer
• When lactose or allolactose is present, it binds with repressor and
inactivates it, allowing RNA polymerase to access to the promoter
region and initiating the transcription
• Regulation by repressor in called negative regulation

Human Genome Project

• Human genome project (HGP) was launched in 1990 to decipher the
complete DNA sequence of the human genome
• Genetic engineering techniques were used to isolate and clone the
DNA segment for determining the DNA sequence
• The project got completed in 2003, the sequence of chromosome 1
was completed in May 2006
• Some of the key findings of HGP are:
 I. The human genome has 3164.7 million bp
II. Total ~30,000 genes are present with an average of 3000
bases per gene
III. . Dystrophin gene is the largest human gene having 2.4
million bases
IV. 99.9 % of nucleotides are the same in all people
V. Only 2 % of genome codes for proteins
VI. Most genes are found on chromosome 1, i.e. 2968
VII. Least genes are found on the Y chromosome, i.e. 231
VIII. There are around 1.4 million locations, where there is a
single base difference in DNA, it is called single nucleotide
polymorphism- SNPs (snips)

DNA Fingerprinting
• Alec Jeffreys initially developed the DNA fingerprinting techniques
and named it as VNTR (Variable Number of Tandem Repeats)
• The difference in the DNA makeup accounts for the unique
phenotype of each individual and that is the basis of DNA
fingerprinting
The DNA sequence of two individuals can be compared very quickly
by the DNA fingerprinting technique
 DNA fingerprinting involves identifying the difference between two
DNA molecule at the specific regions where the sequence is
repeated many times called repetitive DNA
• These repetitive DNAs make a small peak during density gradient
centrifugation and known as satellite DNA
• Depending on the number of repetitive units, base composition and
length of the segment, satellite DNA is further classified into
microsatellites, mini-satellites, etc.
• These sequences do not code for protein but make a large portion of
the human genome
• A high degree of polymorphism present in these sequences is the
basis of DNA fingerprinting
• DNA fingerprinting is used for paternity test as this polymorphism is
inherited to the child
• It has been widely used in forensic science
• DNA fingerprinting can be used in determining genetic diversity
existing in a population

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