Genetics

Genetic material
(RNA & DNA)
GENETIC MATERIAL
 From one generation to the next generation certain substances or
materials get transferred and help the offspring to express
characteristics. This material which gets transferred from parental
generation to the offspring is called genetic material.

 In living organisms, DNA and RNA are the main genetic materials.

 Hence, Genetic materials must be able to:

1. Divide (replicate) and be inherited to the offspring.
2. Carry and process all the information required for the function of
a cell.
3. Change due to mutation.
4. It should have information for the formation of characteristics of
organism
Gene:
 The chemical basis of life is called a gene. It is also the unit
of heredity that is transmitted from one generation to the
another.
 Therefore, a gene is a part or segment of the DNA, which
encodes the character of an organism

GENE (Gr.)= Genesis= to be born

Coined by Danish Genetics- wilhelm Johannsen (1909)
 Properties of Gene
 All genes found in the DNA of an organism which is found on their
chromosomes.
 Each gene is present in a specific position is called a gene locus.

 Genes arrange in linear fashion

 Gene is present in the DNA. DNA is capable of duplication or

multiplication. This is called the process of replication.

 Each gene has a contrasting pair of characters called alleles.

 Alleles must have similar locus in homologous chromosome.

 Genes present in different locus of same or different chromosomes
are called non alleles
 Few genes have more than two alleles called multiple alleles
 Genes are capable of undergoing changes and modification due to
various terminologies like mutation or recombination.
 Functions of genes

Responsible for passing of characters from parents to

offspring

Characters of the organism is determine by genes( it code

for special protein)

Differential expression of genes contained in an

organism’s cell give rise to various specialized cells with
different characters and responsible for different
functions
 Molecular structure of a gene
Chemically, a gene is formed of DNA, hence it may have more than
one functional units. These findings and the foundation of DNA’s
structure and function, Seymour Benzer, in 1962, coined the terms
cistron, muton and recon.

1. Cistron: (Gene as a unit of function) The function unit of the gene

is called cistron. It is also called the genetic unit of DNA molecules
(functionally) because this part of DNA directs the synthesis of a
single polypeptide chain, which controls a single physiological
function. Production of a specific protein is done by cistron.
Hence, to produce one polypeptide or one function, separate
cistron will be present.
2. Muton : (Gene as a unit of Mutation): The smallest part of DNA
which can undergo mutation is called muton. Any change in the
base of a genetic code will modify the message carried by the
codon.
3. Recon: (Gene as a unit of recombination): The smallest
unit of DNA which undergo recombination and crossing
over is called a recon. A recon may be small as one
nucleotide pair of DNA.

A gene consists of only one cistron. A cistron consists of

several recons and a recon consists of several mutons.
Therefore muton is the smallest unit of a cistron.
Central Dogma
The flow of information takes place from DNA to RNA (mRNA) to
proteins. This relation of flow of information is known as central
dogma. Thus it can be said that genetic information flows from
nucleic acids to proteins. The flow of the information is
unidirectional. The concept of central dogma was advanced by
crick in 1958.
Reverse central dogma
 Some viruses commonly known as retroviruses contain RNA as
the genetic material. Hence reverse transcription occurs in such
cases. Reverse transcriptase enzyme plays an important role.
Therefore, information flows from RNA to DNA to mRNA and
finally into protein. Such flow of information is called reverse
central dogma. This was reported by Temin in 1970 and
Baltimore in 1970 in RSV (Raus’s Sarcoma virus, a kind of
retrovirus)
 Nucleic acids
 Nucleic acids are the most essential molecules of the life.

 Nucleic acids are non –protein, nitrogenous substances which

are the polymers of nucleotides.

 These are complex, long chained compounds larger than

proteins.

 They contain C, H, O, N and P.

 First isolated from nucleus of pus by F. Meisher (1869)

 Two types :- DNA (deoxyribonucleic acid) and RNA (ribonucleic

acid).
 Required for the storage and expression of genetic information
 Components of nucleic acids
 Nucleic acids are giant biomolecules made of
monomers called nucleotides (polynucleotides).

 Nucleotides have three components:

I. Pentose sugar (5-carbon sugar),
II. Phosphate group, and
III. Nitrogenous base.
Structure of a nucleotide
 A nucleotide is an organic molecule that is the building
block of DNA and RNA.

 A nucleotide is made up of three parts: a phosphate

group, a 5-carbon sugar (pentose sugar) and
a nitrogenous base.

 It is acidic in nature.

Nitrogenous base + sugar +

phosphate→ Nucleotide
 1. Pentose sugar
 Pentose sugars are the five carbon monosaccharides
found in the nucleic acids.
 RNA = Ribose sugar
 DNA= Deoxyribose sugar
 Ribose and deoxyribose sugar differ in structure at C2.
Deoxyribose has one oxygen less at C2 compared to
ribose sugar.
 2. Nitrogenous bases

 There are two categories of nitogenous bases:

i)Purine Base: Adenine and Guanine
ii)Pyrimidine Base: Thymine, Uracil and Cytosine

i) Purine Base:
 Contains two carbon nitrogen rings in their structure
 Considered as a fusion of pyrimidine with imidazole
ring
 Adenine (A) and Guanine (G)
 Found in both DNA and RNA
ii) Pyrimidine base:
 Pyrimidine have only one carbon nitrogen ring
 Cytosine (C), Thymine (T) and Uracil (U)
 RNA:- U and C
 DNA :- T and C

Complemetary bases
 A purine base is attached to its specific pyrimidine
base with hygrogen bond i.e.,
A=T (two hydrogen bond)
C G (three hydrogen bond)
A G T/U C
Purine
Found
both in
DNA and
RNA
3. Phosphoric acid

 It contains a phosphate group.

 It combines two nucleotides together by

a phospho-diester bond to form a back
bone of nucleic acid.
Nucleoside:

 Nucelosides are formed by combination of purine or

pyrimidine base linked to ᵦ - D-ribose (in RNA) or ᵦ- D-
deoxyribose (in DNA)
Nucleoside: nitrogenous base + sugar
Nucleotide: nucleoside + phosphate
 Nucleotide:
 A nucleotide is a nucleoside with one or more
phosphate group attached to 5’ or 3’ carbon of pentose
sugar
Nucleotide: nucleoside + phosphate
Formation of chain of nucleotides
One nucleotide is linked by another nucleotide by
phosphodiester bond
Nucleotides of each of the two helical strands are bound to
each other by covalent 3’—5’ phosphodiester linkage
The end of the linear strand which bear a free 5’ phosphate
group without phosphodiester linkage is called 5’ end
The opposite end which bears a free 3’ –hydroxyl group is
called the 3’ end
 Deoxyribonucleic Acid (DNA):
 DNA is mainly found in the nucleus of eukaryotic cell, but
also found in mitochondria and chloroplast in little
amount.

 DNA is a polymer of deoxyribonucleotides that contain

many deoxyribonucleotides covalently linked by
phosphodiester bond

 DNA is generally double stranded except in certain

viruses where single stranded DNA is present ( for eg:-
Parovirus)

Those nucleotides are formed by

i) deoxy-ribose sugar ii) Phosphate and iii) nitrogenous base.
 DNA
i) Deoxy-ribose sugar: It is a pentose sugar having 5
carbon atoms. Due to the deoxyribose sugar it is called
deoxyribose nucleic acid.

ii) Phosphate: the phosphate in the DNA is present as

phosphoric acid (H3PO4).

iii) Nitrogenous Bases: the nitrogenous bases are of two

types- Purine and pyrimidine. Purine bases comprise mainly
adenine (A) and guanine (G) while pyrimidine bases
comprise cytosine (C) and thymine (T).
Structure of DNA OR Watson and Crick Model Of DNA
 The three-
dimensional structure of
DNA, first proposed by
James D. Watson and
Francis H. C. Crick in 1953
 Awarded Noble prize in
1962
 Model for DNA known as “
double helix”
 The two DNA strands are
called polynucleotides, as
they are made of simpler
monomer units called
nucleotides
 D.N.A is a polymer of about 10^10 deoxyribonucleotides
 There are only four different types of
deoxyribonucleotides
 Adenine deoxyribonucleotides or Deoxy Adenosine
Monophosphate (d AMP)
 Thymine deoxyribonucleotides or Deoxy Thymidine
Monophosphate (d TMP)
 Guanine deoxyribonucleotides or Deoxy Guanosine
Monophosphate (d GMP)
 Cytosine deoxyribonucleotides or Deoxy Cytosine
Monophosphate (d CMP)
Watson and Crick model of D.N.A
(double helical structure):-
 D.N.A is a polymer of about 10
deoxyribonucleotides per turn.
 D.N.A is a macromolecule made up of
helically twisted two anti-parallel strands.
(One strand moves from 3'-5' direction and
other moves from 5'-3' direction).
 These two strands of D.N.A are called D.N.A
duplex which is spirally twisted around the
central axis in a right-handed manner
 The D.N.A has got many complete turns.
The length of one complete turn is 34°A or
3.4nm and has about 10 nitrogen base
pairs. The distance between two base pairs
is 3.4°A or 0.34nm.
 Each spirally coiled complete turn has one
minor or shallow groove and one deeper or
major groove.
Watson and Crick model of D.N.A (double helical
structure):-
 The strands(backbones) of D.N.A are known as
structural units formed by the combination of
deoxyribose sugar and phosphate groups by
phosphodiester bonds. The distance between two
strands or diameter of D.N.A is 20°A.
 Each deoxyribonucleotide of D.N.A is made up of an
outer phosphate group, middle deoxyribose sugar and
inner nitrogen bases.
 Nitrogen bases are of two types: purine and
pyrimidine. These bases are known as the functional
unit of D.N.A and lie inner side of the helix.
 A purine base is attached to its specific pyrimidine
base. I.e:Adenine combines to Thymine by double
hydrogen bond. ( A=T) and Cytosine combines to
Guanine by triple hydrogen bond. (G≡C)
 A D.N.A looks like a ladder as both strands are joined
together by weak hydrogen bonds in their nitrogen
bases. The total number of purine is equal to the total
number of pyrimidine. (Chargaff's rule). i:e A+G=C+T
 Chargaff’s rule
Chargaff's observation that in the base composition of DNA the
quantity of adenine equal to the quantity of thymine and the
quantity of guanine equal to the quantity of cytosine.

 The amount of A always

equalled the amount of T, and
the amount of C always
equalled the amount of G (A =
T and G = C)
 These findings,
called Chargaff's rules, turned
out to be crucial to Watson and
Crick's model of the DNA
double helix.
 TYPES OF DNA

A- DNA= 11 bp per turn and right handed duplex

B- DNA= 10 bp per turn and right handed duplex
C- DNA= 9 bp per turn and right handed duplex
D- DNA= 8 bp per turn and right handed duplex
Z- DNA= 12 bp per turn and left handed duplex

Two strands
Template strand or sense strand= RNA synthesis
Non template strand or non sense strand= doesnot participate in
RNA synthesis
Circular and linear DNA
 In many prokaryotes , Two ends of a DNA duplex DNA are covalently
linked to form circular DNA. Circular DNA are naked (without
association of histone proteins)
 In eukaryotes the two ends are free to form linear DNA.

Functions of DNA
 It is a Genetic material which carries all the hereditary
information from one generation to other
 It synthesizes ribonucleic acid (RNA) through the process of
transcription
 It acts a prime molecule during protein synthesis
 It controls biological activities of cells. DNA synthesis proteins,
enzymes and other biochemicals
 It guides the process of translation.
 DNA has autocatalytic function which directs the synthesis of
its own carbon copy
 RNA (Ribonucleic acid)
 RNA is a ribonucleic acid that helps in the synthesis of proteins in
our body.

 It is usually obtained from the DNA molecule.

 RNA is a polymer of ribonucleotides of adenine, uracil, cytosine and

guanine which are joined together by phosphodiester bond

 RNA is found in nucleolus, ribosomes, mitochondria, chloroplast and

cytoplasm

 RNA consists of an only single ribose sugar molecule in it.

 Hence is the name Ribonucleic acid.

 RNA is also referred to as an enzyme as it helps in the process of

chemical reactions in the body
 Structure of RNA
A. Primary structure of RNA
 RNA is a single stranded. It consists of
poly ribonucleotides.

 Each RNA consists of a pentose sugar,

phosphate and a nitrogen base.

 RNA has the same nitrogen bases called

the adenine, Guanine, Cytosine as that of
the DNA except for the Thymine which is
replaced by the uracil.

 The nucleotides are joined together by

phosphodiester bonds.

 It forms the backbone of the RNA strand.

B. Secondary structure of RNA
 It involves various coil formation of the polyribonucleotide chain

 These coil structures are stabilized by hydrophobic interaction

between purine and pyrimidine
C. Tertiary structure of RNA
It involves the folding of the molecule into three
dimensional structure
 Functions of RNA
 Facilitate the translation of DNA into proteins.
 Functions as an adapter molecule in protein
synthesis.
 Serves as a messenger between the DNA and the
ribosomes.
 Promotes the ribosomes to choose the right
amino acid which is required in building up of
new proteins in the body.
 In some viruses, it acts as a hereditary material.
Virus such as HIV, TMV etc have RNA as genetic
material
 Types of RNA

 In both prokaryotes and eukaryotes, there are three main types of

RNA (molecular size and function)–

 messenger RNA (mRNA), ribosomal RNA (rRNA), and transfer RNA

(tRNA).
1. Messenger RNA (mRNA):-
 Messenger RNA occurs in the nucleus and synthesis as
heterogenous nuclear RNA (hnRNA) and has linear structure.
 It constitutes about 5% - 10% of the total RNA present in the
cell, m-RNA carries the genetic information from DNA for
Protein synthesis (Translation).
 It act as template for protein synthesis.
 It is short lived and unstable.
Two types:
 Monocistronic: smaller in size because it carries information
from one gene (cistron). Found in eukaryotes.
 Polycistronic: Larger in size because it carries information from a
number of gene (cistron). Found in Prokaryotes.
2. Ribosomal RNA (rRNA)
 it is the most abundant, insoluble and stable type of RNA

 It makes about 70- 80% of the total RNA in the cell. It is the major
component of ribosomal.

 It is synthesized in the nucleus and finally diffuses out in the cytoplasm

and binds with ribosomal protein molecules to form complete
functional ribosomes.

 it play a key role in the binding of mRNA to ribosome during its

translation
 r-DNA is synthesized from the DNA of nucleolus

 Two Types of ribosomes:- 70S (30S + 50S) and 80 S (40S+60S)

3. Transfer RNA (tRNA)
It is about 10-15% of total RNA.

Second most stable type

Smallest RNA with 70-90

nucleotides and is called soluble
RNA.
Each t-RNA serves as an “adaptor”
molecule that carries its specific
amino acids to the site of protein
synthesis
One tRNA carries only one amino
acid.
 Structure of tRNA
 It gets coil over itself to form double stranded
RNA look like a clover leaf (2-D figure).
 Single polynucleotide chain of tRNA is folded
upon itself to form five arms The arms are:-
 The arms are-
1. Acceptor arm:
 Consists of 7 base pairs and 4 unpaired
nucleotide units
 This arm is capped with a sequence CCA (5’ to
3’).
 The amino acid is attached to the acceptor
arm.
 As the amino acid binds to 3’ end of CCA
sequence, hence called as amino acid binding
site.
 The 5’ end bears guanine or cytosine.
2. Anticodon arm:
 This arm is 5-base pair long and a loop called anticodon
loop

 Anticodon loop consists of 7 unpaired nucleotides, 3 of

which are complementary to triplet codon of m-RNA

 It has specific nucleotide bases complementary to the

codons of mRNA which is called anticodon

 The anticodon represents amino acid in tRNA.

3. DHU/ D- arm: It is so named due to
presence of dihydrouridine. It is the binding
site for aminoacyl synthetase enzyme. It is
made up of 8-12 bases. This loop is called
loop I.

4. TψC arm: This arm contains a sequences of

T, pseudo- uridine and C. it has 5 base pairs
and loop consists of 7 unpaired nucleosides. It
is the site for binding tRNA to ribosomes
(ribosome recognition site).

5. Extra/Variable arm: it is a variable site

arm which lies between TψC arm and
anticodon arm. Its exact role is not known. It
may have five bases in its loop.
 Functions of tRNA

 tRNA is necessary at the time of protein synthesis.

 During protein synthesis, a tRNA picks up specific

amino acids from cytoplasm and transport where
protein is to be synthesized. Codons are recognised by
anticodon of tRNAs. Specific amino acids are
recognised by amino acyl synthetase enzymes.
SN DNA RNA

1. It is double stranded , helix and spirally It is single stranded and non helical.
coiled.
2. It has deoxyribose sugar. It has ribose sugar.

3. The nitrogenous bases are A, G, C , T. The nitrogenous bases are A, G, C , U.

4. It is made up of large number of It is made up of small number of nucleotides

nucleotides
5. DNA has equal proportion of purines RNA has no equal proportion of purines and
and pyrimidines. pyrimidines.

6. DNA is of only two types : intra nuclear RNA is three types: mRNA, tRNA and rRNA.
and extranuclear.
7. DNA can replicate itself and form new RNA cannot replicate itself
DNA.
8. DNA is a hereditary material It is not hereditary material except some
viruses eg ribovirus.
9. DNA transcribes genetic information to RNA translates the transcribed message for
RNA. making polypeptides
10. It carries genetic information from RNA participates in protein synthesis.
DNA Replication (Background)
In molecular biology DNA replication is the biological
process of producing two identical replicas of DNA from
one original DNA molecule .

• DNA replication was postulated by Watson and Crick after

they dicoverd the structure of DNA .

• DNA replication is semiconservative . Each srand in the

double helix acts as a template for synthesis of a new ,
complementary .

• The process of DNA replication is vital for cell growth ,

repair , and reproduction in organisms .
What is replication of DNA ?
• DNA replication is the process by which DNA makes
a copy of itself during cell division .

• DNA found within the nucleus , must be replicated

in order to ensure that each new cell receives he
correct number of chromosomes. The process of
duplication is called DNA replication .

• Fundamental process occurring in all cells for

copying DNA to transfer the genetic information to
daughter cells .
Types of replication
1. Conservative replication: The
parental double helix remains intact;
– both strands of the daughter
double helix are newly synthesized

2. Semi- conservative: one strand

from parent in each new strand

3. Dispersive replication: – At
completion, both strands of both
double helices contain both original
and newly synthesized material.
Basic rules of DNA replication
1 . Replication is always semiconservative .
2 . Replication begins at the sequences called origins .
3 . DNA synthesis is initiated by short fragments of RNA call
primers .
4 . The elongation of DNA strands is always in the 5’ to 3’
direction .
5 . DNA replication can be uni or bidirectional .
6 . Replication is continuous on the leading strand and
discontinuous on the lagging strand .
7 . New nucleotide strands are complementary and
antiparallel to their template strands .
8 . Replication takes place at very high rates and is
astonishingly accurate due to the processes of nucleotide
selection , proof reading and repair mechanisms .
Basic requirement for DNA replication
1. DNA helicase:
unwind and separates double stranded as it moves along the DNA .
2. DNA primase :-
A type of RNA polymerase that generates RNA primers . RNA
molecule acts as templates for the starting point of DNA replication.
3. DNA polymerases :-
Synthesize new DNA molecules by adding nucleotides to leading and
lagging DNA strands .
4.DNA Gyrase/ Topoisomerase :-
unwind and rewinds DNA strands to prevent the DNA from
becoming tangled or supercoiled .
5. DNA ligase :-
joins DNA fragments together by forming phosphodiester bonds
between nucleotides .
6.Single strand binding proteins :-
Keep the DNA single stranded after it has been
melted by helicase .
7. RNA primer :-
RNA primer composed of multiple bases that
attached to the template strands to initiate the DNA
replication .
8. Telomerase :-
Finishes off the ends of DNA strands .
9. RNAse:- it is an enzyme which digest the RNA
primers after DNA synthesis is over.
Steps of Replication in DNA
1.Origination of DNA replication

2.Unwinding of parental DNA

3.Initiation of DNA synthesis

4.Chain elongation

5.Excision of RNA primer and their replacement by

DNA

6.Proof reading and DNA repair

1. Origination of DNA replication
 Replication start at specific point of DNA known as
origin of replication (ori).

 In prokaryotes cell, there is one ori where as in

eukaryotic cell number of ori is more than one.

 There is formation of replication bubble due to multiple

origin of DNA replication.

 Replication occurs in both directions along the length of

DNA and both strands are replicated simultaneously.
2. Unwinding of parental DNA
Before DNA can be replicated the double stranded
molecule must be unzipped into two single strands.
 DNA has four base called adenine , thymine , cytosine
and guanine that form pairs between the two strands .
 in order to unwind DNA these interaction between base
pairs must be broken .
 these is performed by an enzyme known as DNA helicase
using energy from adenosine triphosphate (ATP).
 DNA helicase separate the strands into Y shape known as
the replication fork .
 Each seperated strand are stabilized by Single stranded
DNA binding protein (SSB).
 As the two strands are seperated, a problem of positive
supercoil is encountered which is solve by DNA
topoisomerase .
3. Initiation of DNA synthesis

Initiation of DNA synthesis requires primer.

The primer of DNA synthesis is a short

pieceRNA , not DNA .

These short stretches of RNA are produced by

RNA polymerase called primase.
4. Chain elongation
DNA polymerase III elongates a new DNA strand by adding
activated deoxyribonucleotide one at a time to the free 3’
–OH end .

Deoxyribonucleoside monophosphate is activate when

they react with ATP to form deoxyribonucleoside
triphosphate (dATP, dGTP, dTTP,dCTP).

During formation of phosphodiester bond between

sucessive deoxynucleotides, two phosphate groups of
deoxyribonucleotides are removed.

Energy are liberated when phosphate bond is broken

down, thus liberated energy is utilized for the formation of
phosphodiester bond.
DNA polymerases III is involved in DNA replication . Because
replication proceeds in the 5’ to 3’ direction on the leading
strand , the newly formed strands is continuous .

On 5’ to 3’ template there is discountinous formation od

DNA and produces short stretches of DNA which are called
okazaki fragments and the strand is called lagging strands.

The lagging strands begins replication by binding with

multiple primer.

Each primer is only several bases apart .

Okazaki fragments are joined by DNA ligase.

5. Excision of RNA primer and their replacement by DNA

RNA primer is excised by exonuclease activity of DNA

polymerase I and the gap is filled by DNA polymerase I
6. Proof reading and DNA repair

In some cases, there is misinsertion of wrong base. The

probability of misinsetion of wrong base is one in ten
thousand.
Correction is made during proof reading by repair
enzyme.
Significance of DNA replication
DNA replication is necessary for cell division.
DNA replication helps to maintain genetic make
up of an organism.
DNA replication essential for the transmission of
parental characters to offspring.

Leading and lagging strand difference (H.w.)