The genetic material

 DNA –forms the genetic material in most of the

organisms(all eukaryotes, bacteria ,fungi etc)

 RNA – forms the genetic material in some viruses.In

other all organisms it acts as helping factor in the
synthesis of protiens
 DNA
 Is Deoxyribonucleic acid
 RNA is Ribonucleic acid
Structure of DNA
 Is a polynucleotide chain
It has three components
 A) A nitrogeneous base:purines(adenine
,guanine),pyrimidines(thymine and cytosine)

 B)a pentose sugar

 C) a phosphate group
1. pentose sugar
2. NITROGEN BASES
 A nitrogenous base is linked to the pentose sugar by N-glycosidic
linkage
 In DNA, these bases are cytosine (C), thymine (T), adenine (A) and
guanine (G).
 These bases attach in place of the -OH group on the 1' carbon atom in the
sugar ring.
3. PHOSPHATE((phosphoric acid)
 Attaching a phosphate group
 The other repeating part of the DNA backbone is a
phosphate group. A phosphate group is attached to the
sugar molecule in place of the -OH group on the 5'
carbon.This is a phosphoester linkage
 A nitrogeneous base +pentose
sugar=nucleoside( deoxy-
adenosine, deoxy guanosine
guanosine ,deoxycytidine or
cytidine,deoxy thymidine)
 A nucleoside + phosphate
=nucleotide
 A DNA strand is thus made up of
many nucleotides forming a
polynucleotide
 When a phosphate is atached to 5’OH of a nucleoside
through a phosphoester linkage ,a nucleotide is
formed
 Two nucleotides are joined by 3’-5’ phosphodiester
linkage
 Thus a polymer is formed
 A polymer thus formed will be having a free phosphate
at 5’ and free OH at 3’
 Thus a polynucleotide chain has 5’ end and 3’ end (

3’
 Joining the nucleotides into a DNA strand

 A DNA strand is simply a string of nucleotides joined together

A nucleotide
A dinuleotide
A polynucleotide
Watson And Crick’s Model of DNA
 In 1953 James Watson and Francis Crick were awarded nobel prize for
the elucidation of structure of DNA
 They explained the double helical structure of DNA
 Chargaff’s Rule: explains the base pairing of the nitrogenous bases .Acc.
to them the ratio b\w the bases will be constant and will be one .the
purine adenine always base pairs with pyrimidine thymine and guanine
always base pairs with cytosine.
Features of DNA double helix
 Is made of two polynucleotide chains,where the back bone is made of
sugar and phosphate,and the bases project inside
 Two chains have antiparallel polarity ie, if one strand is 5’-3’ direction the
other strand will be 3’-5’
 The base in the strands are paired by hydrogen bonds
 A-T pairing occur by 2-hydrogen bonds and G-C base pairing occur by 3-
hydrogen bonds
 The pitch each of the helix is 3.4nm
 There are almost ten base pairs in each turn
 The distance b\w the base pairs in a helix is 0.34nm
 The plane of one base pair stacks over the other and
this gives stability to helical structure in addition to
the hydrogen bonds
 Length of a DNA is based on the number of nucleotides or on the number
of base pairs
 Lambda bacteriophage has 48052bp , E.coli has 4.6xIO base pairs
 Haploid DNA of humans have 3.3x10 base pairs
 Packaging of DNA
 Human DNA is almost 2.2 meters long
 Such a long DNA is packed with help of Basic proteins called
histones
 Histones are organized to form a unit of eight molecules called
histone octamer
 Histones are positively charged because of arginine and lysine
in the side chains

 the bead like structure found on chromatin formed by

wrapping of negatively charged DNA on the histones is called
nucleosome
 A nucleosome cosist of 200 base pairs
 they appear as beaded structures on the chromatin
LINKER DNA is a double-stranded
DNA in between two nucleosome
cores that, in association with histone
H1, holds the cores together. Linker
DNA is seen as the string in the "beads
and string model", which is made by
using an ionic solution on the
chromatin

Histone octamer H2A H2B H3 &H4x2

 Packaging in prokaryotes
In prokaryotes:
the DNA which lies in the cytoplasm
is held with some proteins which are
positively charged in the nucleoid.

The DNA is organized into loops

which are held by positively charged
protiens
 NHC
 Non histone chromosomal proteins- are the proteins that packs the
chromatin fibers when they attain the shapes of chromosomes
 Euchromatin: loosely packed chromatin
they are transcriptionally active
 Heterochromatin: the part of the chromatin which are densely packed
and which don’t have any role in transcription
SEARCH FOR GENETIC MATERIAL
experimentation
 Transforming Principle of
 The two strains of bacteria were grown in
Griffith
culture medium
 His experiment: he used two
 Both were injected into body of mice
strains of Streptococcus
pneumonia bacteria  S strain bacteria when injected
developed pneumonia and mouse died
 S strain : smooth strained
which develop a  R stain bacteria couldn’t cause
polysaccharide coat around it pneumonia and mouse survived
 R strain: rough strain where
the coating is absent
2nd step Conclusion
 He heat killed S strain bacteria and  In mixture some factor present in heat
injected to mice and mouse didn’t killed S strain has transformed R strain
develop pneumonia into virulent form
 The R strain thus produced mucous coat
3 rd step
and became infectious

 A mixture of heat killed S strain and living R

strain were injected mouse died of
pneumonia
AVERY AND MAC CARTY ‘S EXPERIMENT TO PROVE TRANSFORMING PRINCIPLE
Hershey chase’s experiment to prove DNA is the genetic
material
Aim :to identify whether Dna or protein is  Similarly viruses grown in
the genetic material radioactive phosphorous were
 They grew some bacteriophages in two found to be having
different medium one containing radioactiveDNA and their
radioactive sulphur and some in proteins were not radioactive
radioactive phosphorous
 Observation: they found that those
viruses grown in medium with
radioactive sulphur had their
proteins which became radioactive
but their DNA was not radioactive
2nd step
 Bacteriophages were attached
toE.coli bacteria to develop
infection
 After infection the viral coats were
removed by agitating a in a blender
 Centrifugation was done to to
separate viral particles from
bacteria
 RESULT: The bacteria which were infected
with viruses that had radio active DNA were
radioactive but those bacetria which were
infected by viruses with Radioactive
Proteins were not radioactive
 This indicates that the protiens have not
entered the bacteria proving that DNA is
the genetic material
Replication of DNA
 Occurs in the S phase of the cell cycle
 Watson and Crick explained the semi-conservative mode of DNA replication
 Acc to them when Dna replicates the two strands separate .(both act as
parental or template strands)
 A complimentary strand is produced for each parental strand
 Thus after replication each DNA will have one parental strand and one
daughter strand
Semi conservative mode of DNA replication
Experimental proof for semiconservative mode of replication
 Meselson and Stahl’s experiment
 They grew E.coli bacteria in a medium containing 15NH4Cl(heavy isotope of N2)for
several generations
 They observed that the DNA and nitrogenous compounds in the bacteria possessed
15 N
 They observed DNA by centrifuging in cesium chloride
 Cesium chloride forms a concentration/density gradient when used with nucleic
acids in a centrifuge. It allows separation of nucleic acids based on their weights. The
heavier nucleic acids or pieces go toward the bottom of the cuvette.
 Later they transferred these cells into normal 14NH4Cl and observed the DNA
at various intervals
 The DNA obtained after 20minutes was a hybrid DNA which was of
intermediate density
 After 40 minutes it was observed that cuvette had equal number of hybrid and
light DNA
Process of replication
 Longer DNA molecules will not replicate together
 Replication starts with binding of an enzyme helicase which
helps in unwinding the strands
 The opening formed by the breaking of hydrogen bonds by
helicases where replication occur is called replication fork
 The strands separated are called parental strands
 On the parental strand which is in 3’-5’ direction DNA
polymerase enzyme initiates the synthesis of complimentary
strand . The formation of daughter strand in this case is
continious ,it is called as leading strand
 On the parental strand which is in 5’-3 direction the DNA polymerase cannot initiate
synthesis of complimentary strand
 Here an enzyme primase produces an RNA primer
 The RNA primer will produce the daughter strand in 3’-5’ direction
 Here a new strand is produced discontinously and is called lagging strand .
 The fragments formed are called okazaki fragments
 The newly formed daughter strand will be joined by an enzyme called ligases
 DNA polymerase: is the enzyme that catalyses polymerisation of
nucleotides DNA polymerase is functional in 5’-3’direction only
 Deoxy ribonucleoside triphosphate provide energy required for DNA
replication
 They act as substrates for Dna replication
 In E.coli the replication orginates from orgin of replication only
 RNA
DNA

 Is ribonucleic acid
 Nucleotides will have an additional
OH group at 2’position
 It has uracil at the place of thymine in DNA

RNA
IDEAL Genetic material
 Should be able to replicate
 Should be structurally and chemically stable
 It should provide scope for slow mutations needed for evolution
 Should be able to express itself in the form of Menedelian characters
RNA WORLD
 RNA is the first genetic material
a)essential life processes like metabolism splicing translation evolved
around RNA
b)RNA is the genetic material for some viruses
c)RNA is reactive and catalayse reaction
WHY DNA better Genetic material
 RNA is highly unstable and reactive due to 2’OH group and mutates
fastly(makes RNA labile and easily degradable)
 DNA is evolved from RNA with chemical modifications that make it more
stable
 DNA is double stranded(RNA is single stranded)and having
complementary strand (Presence of H- bonds and stacking of
Bases)increases its stability
Types of RNA
 mRNA: is an RNA produced by the DNA on which the genetic
information will be copied .the mRNA comes out of the nucleus
 tRNA :transfer RNA , it helps to read the information on mRNA and
collects amino acids from cytoplasm for transalation
 rRNA -ribosomal RNA which synthesies protien chains
 DNA  RNA
 Has deoxy ribose sugar  Has ribose sugar
 5-methyl uracil(thymine) is present  Has uracil in the place of thymine
 Mostly DNA act as genetic material  RNA act as messenger and adaptor molecule (genetic
 DNA is stable material in some viruses)
 Due to 2’OH at every nucleotide makes RNA more labile
and easily biodegradable

 Mutation rate is faster

 Chemically less reactive ,mutate slowly

 RNA directly codes for Proteins

 DNA require RNA for protein synthesis
Central dogma of molecular biology
replicate

transcription

transalation
 protien synthesis(gene
expression)
 It involves two steps :transcription and translation
 Transcription is copying information's present on DNA into an RNA
 Transalation is sequential arrangement of amino acids to form a protein
based on the information present on mRNA
 transcription
 Is the process of producing a complimentary copy on mRNA from DNA
 the strand of DNA on which transcription occur is called template strand
..this strand will be in 3’-5’ direction
 The complementary strand of the template strand is called coding strand
gene
 Is the functional unit of inheritance
 Gene coding for a protein is called cistron
 eukaryotes are monocistronic: dna segment coding for a polypeptide
 prokaryotes are polycistronic
 Eukaryotes shows split gene arrangement ,ie b\w the coding sequences
there will be non coding sequences
Transcription unit

Structural genes
A transcription unit consist of :
 A promoter:is a dna sequence
on which the attachment of
RNA polymerase occur
,transcription starts from
promoter
 Structural genes: the dna
sequence on which
transcription occur
 Terminator:the dna sequence
which determine where the
transcription must stop
 ALL the references for the
transcription is made with
coding strand
 RNA synthesis involves separation
of the DNA strands and synthesis
of an RNA molecule in the 5' to 3'
direction by RNA polymerase,
using one of the DNA strands as a
template.
 In complementary base pairing, A, T,
G, and C on the template DNA strand
specify U, A, C, and G, respectively,
on the RNA strand being
synthesized.
Need of processing in eukaryotes
 In the mono-cistronic DNA which gets transcribed to RNA there are
certain sequences which are non coding for amino acids .these are called
as introns
 They will be seen along with exons (coding sequences)
 a) these introns has to be removed

 RNA polymerase I synthesizes a precursor of rRNA 45S (35S in yeast),
which matures into 28S, 18S and 5.8S rRNAs which will form the major
RNA sections of the ribosome.
 RNA polymerase II synthesizes precursors of mRNAs(hnRNA) . This is the
most studied type, and, due to the high level of control required over
transcription, a range of transcription factors are required for its binding
to promoters.
 RNA polymerase III synthesizes tRNAs, and other small RNAs found in
the nucleus (snRNA)and cytosol (5srRNA).
Genetic code
 There are 4 bases ATGC which has to code for all the 20 amino acids
required by our body
 George Gamow proposed that for 4 bases to code for 20 amino acids the
amino acids should be coded by at least 3 bases
 Genetic code provide a dictionary for translating the language of RNA into
language of proteins
 There are thus 64 codons in a genetic code
GENETIC CODE
FEATURES OF GENETIC CODE
 The codon is triplet.
 There are are 61 codons which codes for different amino acids and the
remaining 3codons are called stop codons which do not code for any
amino acids and they determine the stopping of
transalation(UAA,UAG,UGA)
 One codon codes only for one aminoacid therefore it is said to be
specific and unambigious
 Some amino acids are coded by more than one codon therefore it is
said to be degenerate
 The codon is read on mRNA in a continious fashion without any
punctuations
 The code is said to be universal since codons like UUU in bacteria and
humans code for phenylalanine (phe)
 AUG codon has dual function as I t codes for methionine and is also the
start codon


t RNA
 It reads the code present on the mRNA and links with the aminoacids
 They carry aminoacids to ribosomes where they are linked into protiens.
 they act as an adaptor molecule as it binds with specific aminoacids
 There are specific tRNA,s for each aminoacid
Structure(clover leaf)

 Has 4 regions
 A)anticodon site : this site has
complimentary bases for the codons on
mRNA
 B)enzyme site:or amino acid synthetase
recognition site
 C)ribosome recognition site
 D) amino acid binding site
a mRNA
 UTR,s
:untransalated
regions are the
fragments present in
an mRNA which are
essential for
efficient transalation
ribosomes
 ribosomes are the organelle (in all cells) where proteins are synthesized.
They consist of two-thirds rRNA and one-third protein. Ribosomes consist
of a small (in E. coli , 30S) and larger (50S) subunits. The length of rRNA
differs in each.
 The smaller subunit has a binding site for the mRNA. The larger subunit
has two binding sites for tRNA.ie A -site andP –site

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tRNAs bring their amino acids to the
open binding site on the
ribosome/mRNA complex, forming
a peptide bond between the amino
acids
 transalation ii), the large subunit joins the
Is the polymerisation of small one to form a complete
aminoacids to form a ribosome the larger sub unit
polypeptide attaches to the AUG codon on
 The order and sequence of aminoacids
mRNA and the smaller subunit to
are determined by sequence of base on
the initiator tRNA with amino
mRNA .it involves
acid methionine the protein
synthesis is initiated.
 A)initiation”
 i)the amino acids present in the
cytoplasm bind with ATP and aminoacyl
RNA synthetase enzyme which is specific
to each aminoacid and this is transferred
to tRNA.this process is called charging
of tRNA
 Elongation: A new charged
tRNA enters , at the next codon
downstream of the AUG codon.
If its anticodon matches the
mRNA codon it basepairs and
the ribosome can link the two
aminoacids together. The
ribosome then moves one
triplet forward and a new
tRNA+amino acid can enter the
ribosome and the procedure is
repeated.
 Termination: When the
ribosome reaches one of three
stop codons, for example UGA,
there are no corresponding
tRNAs to that sequence.

 At the stop codons releasing

factors binds and the ribosome
dissociates from the mRNA.
When the ribosome is released
from the mRNA, its large and
small subunit dissociate. Thus
the polpepttide chain is released
 regulation of gene expression
 the formation of polypeptide chain is regulated at
-a)Transcriptional level
- b) Processing level
- c)Transport of mRNA from nucleus to the cytoplasm
-d)Translational level
 Prokaryotic gene expression: mainly at transcriptional level
 The activity of Rna polymerase enzyme is regulated by
some accessory proteins which affects its ability to
recognise the start sites
 Thus the accessibility of promoter is regulated by the
interaction of proteins with sequences termed as operators
 The proteins can acts as inducers or repressors
Lac operon –Jacque Monad
 A polycistronic structural gene is regulated by a common promoter and
regulator gene;such an arrangement is called operon
LAC OPERON:OPERON REGULATING LACTOSE METABOLISM
 an enzyme called beta galactosidase is essential for the break down of
lactose into galactose and glucose that act as a energy source for the E.coli
bacteria
 If the bacteria do not have lactose around them the enzyme will not be
produced
 In lac operon a polycistronic
structural gene is regulated
by a common promoter and
regulatory gene this
arrangement is called operon
 It consist of one regulatory
gene –i gene that codes for
the repressor protein for the
lac operon
 The three structural genes are
z y and a
 The z gene codes for enzyme
beta galactosidase
 The y gene codes for
permease(increase
permeability of the cell to b-
galactosidase) and a gene for
trans-acetylase
 Lactose acts as an inducer
 Condition I: if lactose
absent
 i gene codes for
repressor mRNA and form
the repressor protien ,the
repressor protien binds
with the operator and
prevents RNA polymerase
from transcribing the
structural genes ie.z y and
a genes
 Condition 2:if lactose present
 i gene codes for the
repressor protien but since
the inducer lactose is present
repressor cannot bind with
the operator
 thus operator is free
and allows the polymerase
enzyme to bind with
promoter leading to
transcription of structural
genes
 Thus regulation of lac operon
is considered to be a negative
regulation
DNA fingerprinting
 Helps in identifying differences in some specific
regions of DNA called as repetitive DNA
 Also called as satellite DNA
 They do not code for any protiens
 They form a large part of human genome
 They shows polymorphism
 DNA polymorphism forms the basic principle behind
fingerprinting
 Since this polymorphism is exhibited in same degree
in all tissues like saliva blood bone follicles sperms etc
,they act as tools of forensic studies
 This polymorphism is inherited therefore DNA finger printing can be
used to solve paternal issues
 DNA poymorphism is mutation that are inheritable
 In DNA finger printing DNA polymorphim of satellite sequences are
studied