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Second year
Chapter 6

Chromosomes and DNA

A thread-like structure of nucleic acids and protein found in the nucleus of most
living cells, which can carry genetic information in the form of genes called
chromosome.

Things to be study
 Chromosome
 Types of chromosomes
 Chemical composition of chromosome
 Ultra-structure
 Chromosome as carrier of gene
 Chromosomal theory
 DNA as a hereditary material
 DNA structure
 Watson and crick model of DNA
 Replication of DNA
 Gene the unit of hereditary information
 One gene one enzyme hypothesis
 Cell use RNA to make protein
 Gene expression (Transcription and Translation)
 Genetic code
 Mutation (Example of Gene mutation)
 DNA damage
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Chromosome means colored body and the name given to chromosome by W.

Waldeyer (1888) is wrong because it is not a colored body. Walther Flemming had
discovered chromosome in 1882 while he was examining the larvae of salamander.
He dyed the larvae with Perkin’s Aniline and observed the slightly darker structure
than salamander’s larvae and he named it as chromosome.

We can define Chromosome as a thread-like structure of nucleic acids and protein

found in the nucleus of most living cells, carrying genetic information in the form of
genes and present in a pair in an individual.

The number of chromosome is constant however it varies from species to species. Such as,

Species No. of Chromosomes

Penicillium 2 chromosomes—01 pair
Mosquito 6 chromosomes—03 pairs

Drosophila 8 chromosomes—04 pairs

Garden pea 14 chromosomes—07pairs
Frog 26 chromosomes—13 pairs

Human 46 chromosomes—23 pairs

Sugar cane 80 chromosomes—40 pairs
Fern 1000 chromosomes—500 pairs

Each chromosome consists of two thin threads called chromatids. Each chromosome
has two identical strands called sister chromatids. Chromatids are attached to each
other by a common point called centromere. Within the centromere a disc shaped
structure is present called kinetochore. Kinetochore is the site to which the spindle
fiber attached during cell division. At the beginning of leptotene stage (the first stage of the
prophase-I of meiosis, during which each chromosome becomes visible as two fine threads, the
chromatids) the chromosome seen as an extremely thin thread called chromonema (the
spirally coiled central filament of a chromatid along which the chromomeres are united). Along
chromonema chromomeres are present which are beadlike granules occurring in
series along a chromonema. The length of the chromosome from its centromere to the
terminal end is called arm of the chromosome.

Types of chromosomes
Based on position of centromere

i- Metacentric
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A type of chromosome having the centromere in the median position so that the arms are of
equal length is called metacentric chromosome.

ii- Sub-Metacentric

A type of chromosome having the centromere situated so that one chromosome arm is
somewhat shorter than the other called sub metacentric.

iii- Telocentric

A type of chromosome having the centromere is in a terminal position called telocentric

chromosome.

iv- sub-telocentric or Acrocentric

A type of chromosome having centromere at sub terminal position so that one arm is much
longer than the other called sub-telocentric or acrocentric chromosome.

Based on functions

i- Autosomes

A type of chromosome which are similar in males and females of same species are
called autosome.

ii- Allosomes (Sex chromosome)

A type of chromosome which are different in males and females of same species are
called sex chromosomes. Chromosomes are present in pairs and in this pairing, they
are named as X and Y chromosomes. Male possess XY and female possess XX
chromosomes, while in some animals this position is reversed, in them male possess
like chromosomes and female possess unlike chromosomes e.g. Moth, birds and some
fishes etc.

In these animals, sex-chromosomes are referred by Z and W instead of X and Y so, in

this way female possess ZW and male possess ZZ genetic constitution.

Karyotype
The orderly arrangement of chromosome that an individual possess is called Karyotype.
Chromosomes are paired according to matching, banding and arranged by size and
shape. The karyotype of individual is often examined to detect the genetic abnormalities
that arise from chromosome numbers.

Chemical composition of chromosomes

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Chemically chromosome is made up of deoxy-ribo-nucleoprotein, means DNA and

protein. The chromosomal protein is called histone. DNA is the polymer of nucleotide
and each nucleotide is made of three sub units;

 5-Carbon monosaccharide (i.e. pentose sugar)

 Nitrogenous base
 Phosphoric acid

Ultra-structure of chromosome
Chromosomes are composed of chromatin which is a complex of DNA and protein. The
eukaryotic chromosome is 60% protein and 40% DNA. RNA also associated with this.
The DNA is very long double stranded fiber known as Duplex, it is more than 7 feet (2
meter) in length if laid out from nucleus in a straight line.

It resembles a string of beads. The DNA duplex (of 200 nucleotides) is coiled around a
complex of histones protein, which are small basic polypeptides having positive
charge, and rich in amino acids Arginine and Lysine. Eight of these histones form the
core body for the assembly of DNA duplex called a nucleosome, because so many of
their amino acids are basic, histones are positively charged. The DNA duplex which is
negatively charged is strongly attracted to the histones and wrap tightly around the
histone core i.e. nucleosome.

The core thus acts to promotes and guides the coiling of the DNA. Further coiling of the
DNA occurs when the string of nucleosomes wraps up into higher order coils called
super coils. Chromosome has two parts hetero chromatin and euchromatin.

Heterochromatin
 More condensed
 Silenced genes
 Gene poor (high AT contents)
 Stain darker

Euchromatin
 Less condensed
 Gene expressing
 Gene rich (high GC contents)
 Stain lighter

Chromosomes as carriers of genes

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Genes are information molecules, carry genetic information and play vital role in
inheritance. Genes are located on chromosomes. Chromosomes carry genes to new
offspring. In genotype one member of each paired gene is contributed by one parent
and other by the other parent similarly chromosomes as by the fusion of male and
female gamete zygote is formed. Chromosomes are present in pairs, these are called
homologous chromosomes, each of which contributed by mother and father.
Homologous chromosome has homologous allele (an opposite and changed form of
gene). Genes are segregated during gametes formation.

CHROMOSOMAL THEORY OF HEREDITARY

Chromosomes are vehicles for the information of hereditary. Chromosomal theory of
inheritance was formulated by Walter Sutton and Theodor Boveri in 1902.

Boveri and Sutton's chromosome theory of inheritance states that genes are found at
specific locations on chromosomes, and that the behavior of chromosomes during
meiosis can explain Mendel’s laws of inheritance.

Points of chromosomal theory

i. Chromosomes are present in pairs and known as homologous chromosomes.
ii. Homologous chromosomes segregate during meiosis.
iii. Homologous chromosomes assort themselves independently.

Objection
The objection on chromosomal theory of hereditary is that the number of factors (genes)
is more than the number of chromosomes.

Evidence of DNA as Hereditary Material

DNA possess transforming ability and it transform hereditary characters from one
generation to the next generation. The DNA is found on chromosomes. There are two
theories which explain this transforming ability of chromosome.

1) Fred Griffith’s experiment

Fred Griffith selected two varieties of bacteria for his experiment.

 Rough surface bacteria (nonvirulent and non-capsulated)

 Smooth surface bacteria (virulent and capsulated)

Steps of experiment
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 In first step Griffith injected rough surface bacteria into mice he observed no
effects and mice remained alive.
 In second step Griffith injected smooth surface bacteria into mice he observed
mice died.
 In third step he changed the virulent type of bacteria into non virulent type by
heating them and then injected into the mice, mice remained alive.
 In fourth step he injected dead Smooth surface bacteria along with non-virulent
rough surface bacteria he observed that mice died.

By the Griffith experiment it is proved that the genetic property of dead smooth virulent
bacteria was transformed into rough non-virulent bacteria and converted them into
virulent type.

2) Hershey and chase experiment

Hershey and Chase proved DNA as genetic material. They performed their experiment
on Bacteriophage virus. They labeled Bacteriophage DNA with radioactive p32 and
protein coat with s35. This Bacteriophage allows to attack on bacteria. When the
bacterial cells were checked the p32 found only. It shows that DNA function as genetic
material.

STRUCTURE OF DNA
The DNA is the polymer of nucleotide. Nucleotide is made up of three sub units.

 Pentose sugar
 Nitrogenous base
 Phosphate group (PO4-)

The pentose sugar is deoxy ribose in DNA and ribose in RNA, the nitrogenous bases
are of two types.

1) Purines (double ringed): ---> These are adenine (A) and guanine (G).

2) Pyrimidines (single ringed): ---> These are cytocine (C) thymine (T) and uracil (U).

The nitrogenous base combines at first carbon atom of deoxy ribose to form nucleoside
and phosphoric acid combines with deoxy ribose at its 5th carbon atom to form
nucleotide. These nucleotides are joined with one another through phosphoric acid in
such a way that c3 position of one deoxy ribose linked with c5 position of the next.

WATSON AND CRICK MODEL OF DNA

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Watson and Crick studied the structure of DNA and proposed its double helix model.
According to the Watson and Crick model the structure of DNA molecule is as follows.

i. Each molecule of DNA is made up of two polynucleotide chains, which are

twisted around each other and form a double helix (screw like).
ii. On both side of this helix sugar and phosphate group are present.
iii. Nitrogenous bases are found in the middle of helix. Adenine is always attached
to Thymine and Guanine is always attached to Cytosine.
iv. Double strand of DNA attached to each other with weak hydrogen bond. There
are two hydrogen bonds (=) between Adenine and Thymine and three hydrogen
bonds (≡) between Guanine and Cytosine.
v. Between the two strands of DNA there is 20 A0 distance.
vi. The coiling of double helix is right handed, and complete turns occurs after 34 A0.
vii. Each nucleotide covers 3.4 A0 distance so, 10 mono-nucleotides are found per
complete turn.

Chargaff’s rule
 One strand of DNA has Adenine the other one has Thiamine, similarly, one
strand of DNA has Guanine the other one has Cytosine.
 The amount of Adenine is equal to Thymine and the amount of Guanine is equal
to Cytosine.
 The concentration of purines is equal to concentration of pyrimidines.

The combinations of nitrogenous bases don’t restrict the sequence of nucleotide along
each DNA strand. Thus, the linear sequence of the four bases can vary in countless
ways, and each gene has a unique order or base sequence.

REPLICATION OF DNA
The formation of new DNA molecule is known as replication of DNA. The DNA
polypeptide chain has an imaginable base sequence, with the help of this sequence we
can determine its partner in the duplex for example, if the sequence of one chain is
ATTGCAT, then the sequence of its partner strand must have TAACGTA it shows
complete mirror image of the chain.

The process of DNA replication is as follow;

 The DNA molecule is made up of two chains of polynucleotide which are coiled in
the form of spring.
 Between these two chains weak hydrogen bonds are present. When these
hydrogen bonds destroy the two strands separate like two parts of zip with the
help of Helicase enzyme and form replication fork.
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 Due to this separation the nitrogenous bases like Purines and Pyrimidines
separated from each other.
 In the nucleus of cell already prepared free nucleotides are present; they are
attracted towards the separated Purines and Pyrimidines.
 An Adenine is attached to Thymine and Guanine is attached to Cytosine with a
new H-bond.
 An enzyme Primase starts DNA replication; this enzyme makes a small piece of
RNA called Primer which is the starting point.
 An enzyme Polymerase binds to primer and start the new strand of DNA.
 DNA polymerase can add bases in one direction from the 5-prime end to the 3-
prime end.
 One of the new strands of DNA is leading strand is made continuously.
 Polymerase adding bases one by one.
 The other strand is lagging strand can’t make in this continuous way because it
runs in opposite direction.
 DNA polymerase make it in a series of small chunk called Okazaki fragments.
 Each fragment started with RNA primer from 5 to 3 prime end.
 An enzyme Exonuclease removes all the RNA primers from both strands.
 Another DNA polymerase fill the gap that are left behind with DNA.
 Finally, DNA Ligase seals up the fragments of DNA in both strands to form a
continuous double strand.
 When nucleotides are attached, then sugar of one nucleotide is attached to the
phosphate group of another nucleotide.
 At this time the old DNA coil is completely de-coiled, and its two strands are
separated from each other.
 Each strand is linked to the part of new nucleotide and two new strands are
formed again. They are coiled together in the form of a double helix.

REPLICATION IS SEMI CONSERVATIVE

According to Watson and crick model the replication of DNA molecule is semi conservative.

Semi conservative
The normal process of DNA synthesis, in which original strands of DNA molecule
separate and each act as template on which a new complementary strand is laid down.
In semi conservative replication the original duplex of the DNA is not conserved;
instead, each becomes part of another duplex. It indicates that half part of the DNA
remains in its original shape.
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The complementary nature of the DNA duplex provides the basis of duplicating the
molecule. When two strands of DNA are separated from each other the already
prepared nucleotides are attached to each strand. In this way similar type of two
daughter duplexes of the same sequence are formed.

MATHEW MESELSON AND FRANK STAHL EXPERIMENT

Replication of DNA is semi conservative was tested by Mathew and Frank (1958) in
California institute of technology.

They grew Escherichia coli bacteria for several generations in ammonium chloride
(NH4Cl) medium containing the heavy isotope of nitrogen 15N. When DNA is extracted
from these cells the DNA was found denser than normal. After that E. coli cells with only
15N in their DNA were transferred to a new medium containing lighter isotope 14N and

were allowed to divide. DNA was extracted at various level and dissolved in cesium
chloride (CsCl) and centrifuged thus, DNA strands of different densities got separated.

 A DNA molecule in which both strands are heavy (N15 is most dense)
 After one replication the DNA was found to have intermediate density i.e. a
hybrid DNA molecule in which one strand is heavy and one is light.
 A DNA molecule in which both strands are lighter (N14 is least dense)
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It shows that after one round of replication, each daughter DNA duplex possesses the
one heavy strand and one normal strand. When this hybrid duplex replicated it forms
one heavy strand to form another hybrid duplex and one light strand to form a light
duplex. Thus, this experiment clearly confirmed that DNA replicates in semi
conservative manner.

GENE
The fundamental, physical, and functional unit of heredity is called gene. Gene controls
the specific cellular functions either by synthesizing enzymes, proteins or by regulating
the action of other genes.

British physician Archibald Garrod and William Bateson examined the appearance of
diseases for several generations in certain families. They have noted that some
diseases are more common in some patient’s families. They have observed that genes
are responsible to transfer the hereditary information into new generation.

Alkaptonuria disease
The Alkaptonuria patient’s urine turned black on exposure to air. Such urine contains
homogentisic acid (HGA or alkapton), produced in the liver from excess amount of
tyrosine rapidly oxidized when exposed to air. In normal person homogentisic acid
breaks into simple substances such as CO2 and H2O with the help of HGA oxidase
enzymes. But the affected person lacks this enzyme and homogentisic acid not broken
down into simple compounds. This lack of enzyme character is controlled by autosomal
recessive gene. In early life there is no apparent ill-effect but later on alkapton is
deposited in cartilage that’s why the tip of the nose and pinnae of the ears turns black. A
painful form of arthritis often accompanies.

GENOME
Full set of genes or full sets of chromosomes in each cell is called genome. Example;
single circular chromosome in bacterial cell is genome of bacteria, twenty-two pairs of
autosomes along with a pair of sex chromosomes in human constitute genome.

THE ONE GENE-ONE ENZYME HYPOTHESIS

One gene is responsible for the function of a particular enzyme though this concept is
not accepted nowadays because we know that sometimes multiple genes are required
to produce many different proteins. Beadle and Tatum first proved it by experiments that
each gene encodes the structure of a single enzyme. They have selected bread mold
Neurospora crassa for their experiment.
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Neurospora crassa
Neurospora is the genus of ascomycetes fungi, it is also called red bread mold because
it grows on bread and appear red in color. It is a saprophytic fungus and obtains its food
from bread. Neurospora can easily be grown in laboratory in an artificial medium
containing salt (NaCl), carbon source (glucose and fructose) and vitamin (Arginine or
Thiamine etc.).

Mutation of Mold
Beadle and Tatum forced mutation in ascospores by treating them with X-rays so,
some of the spores became fragile and they cannot grow in the absence of particular
type of vitamin and these types of mutant are called auxotroph. These mutant spores
did not show growth in incomplete medium. In this way, they had experienced damage
to DNA in a region encoding the information of proteins that Neurospora needs for
normal growth such as vitamins.

Mutation: --- the process of alteration or change.

Mutant: ----- an individual or organism that is altered or changed in mutation.

Identifying mutant series

For the identification of mutant genes Beadle and Tatum transferred the spores of
Neurospora into test tubes containing different mediums such as complete medium or
incomplete medium. Complete medium contains salt (NaCl), carbon source (glucose
and fructose) and vitamin (Arginine or Thiamin), incomplete medium contains glucose
and salt or glucose and vitamin. The auxotroph start growth in whole medium and in
glucose and vitamin medium but not in glucose and salt medium. When they added
amino acids such as Ornithine, Citrulline, Arginine to each test tube the mold starts to
grow and then identified the nature of amino acid which was responsible for growth. In
this way Beadle and Tatum develop three different mutant strains of Neurospora and
succeeded in the identification and isolation of many mutant deficient.

 Mutant strain 1: would grow if O, C, or A were added to the minimal medium.

 Mutant strain 2: would grow if either C or A was added to the minimal medium.
 Mutant strain 3: would grow if only A was added to the minimal medium.

Beadle and Tatum concluded that each mutation alter a single gene that controls
the synthesis of particular kind of molecule.

Some prior substances Gene 1 Gene 2 Gene 3

Acetylcholine O C A
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It shows that gene 1 function as to convert some prior substances into O and gene 2
convert O into C and gene 3 convert C into A. If gene 1 is damaged the mold would no
longer grow unless its normal function of synthesizing O was corrected so, adding O to
minimal medium compensates or corrects the loss of gene 1. It is noted that by adding
C or A can also overcome this deficiency.

When they examined these mutations on chromosomes and found these mutations on
three places, named as three gene clusters.

Gene 1 cluster: in gene 1 cluster enzyme E damaged which can convert acetylcholine
compound into ornithine

Gene 2 cluster: in gene 2 cluster enzyme F damaged which can convert ornithine into citrulline

Gene 3 cluster: in gene 3 cluster enzyme G and H damaged which can convert citrulline into
arginosuccinate and arginosuccinate into Arginine.

By this experiment Beadle and Tatum concluded that a defect in one enzyme is due to one
gene present on chromosome. It shows that each gene encodes the structure of a single
enzyme, it is called one-gene one-enzyme hypothesis.

Cells use RNA to make protein

As we know that DNA gives instruction for the synthesis of particular type of protein.
These instructions transferred from the nucleoplasm to cytoplasm with the help of RNA.

To check out that cell uses the RNA for protein synthesis placed the cell in a medium
containing radioactive amino acids. As the proteins are made up of amino acids the cell
will take up these amino acids labeled with radioactive for protein synthesis. This is
known as pulse labeling. So the radioactive proteins were first appeared in the
cytoplasm on ribosome rather than nucleus. Ribosomes are the sites of protein
synthesis. In the protein synthesis three types of RNA take part.

i) Ribosomal RNA (rRNA)

Ribosomal RNA found attached to ribosomes during protein synthesis. It provides site
for the amino acid assembling on ribosomes.

ii) Transfer RNA (tRNA)

Transfer RNA is a small RNA that transport amino acids from cytoplasm to ribosomes
for protein synthesis and rout each amino acid to correct position on polypeptide chain.

iii) Messenger RNA (mRNA)

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Messenger RNA molecule is a long single strand of RNA that passes from the nucleus
to the cytoplasm. Messenger RNA brings information from the chromosome to ribosome
to order the assembly of amino acid of the required protein for polypeptide chain.

AN OVERVIEW OF GENE EXPRESSION

All the functions in the body of an organism are controlled by genes so, a function
expressed or performed by a gene is called gene expression.

In gene expression an RNA copy of each active gene is made which gives instruction of
protein synthesis. The mechanism of protein synthesis is similar in prokaryotes and
eukaryotes because all organisms have a similar trend of gene expression. The
mechanism of gene expression occurs in two phases i.e. transcription and translation.

i- Transcription
The production of RNA from DNA is called transcription. In this process genetic
information are transferred from DNA to RNA. The process of transcription is completed
in three steps.

Step 1: Initiation phase

RNA polymerase binds to a particular sequence of nucleotides or promoter region.

How RNA polymerase knows the promotor region? The answer is that, before promotor
region in prokaryotes -10 TATAAT sequence and -35 TTGACA sequence present while
in eukaryotes -25 TATA sequence and -70 CAAT sequence present. These sequences
are present before the initiation site of gene.

When RNA polymerase binds to DNA template strand or antisense strand a

transcription bubble is appeared.
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Step 2: Elongation phase

In protein synthesis pathway elongation phase is responsible for the growth of nascent
polypeptide chains or RNA.

The RNA polymerase starts the synthesis of RNA strand with a nucleotide sequence
complimentary to DNA strand by moving on DNA strand, as the RNA polymerase start
moving on DNA strand meet different DNA nucleotides sequence and adds related
similar RNA nucleotide to the newly form RNA strand.

Step 3: Termination phase

At the terminal end of DNA strand stop signal is present such as series of GC base
pairs followed by AT base pairs. When polymerase enzyme reaches at stop signal, it
separates from the DNA and forms GC hair pin in RNA molecule. RNA is then
transported into cytoplasm where it takes part in protein synthesis. This RNA is known
as messenger RNA and it contains a message or code in the form of a triplet of three
nitrogen bases it is called codon triplet.

Post transcription changes in RNA

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RNA molecule has many enemies specially enzymes that can hydrolyzed RNA
molecule. To minimize the effect of enzymes, RNA develop caps on their 5 prime and 3
prime end. On 5 prime end 7-methyl Guanosine Tri Phosphate cap is formed while on 3
prime poly Adenine cap is formed.

ii- Translation
The process of polypeptide chain synthesis on ribosome under the direction of mRNA is
called translation or conversion of nucleotide sequence of DNA into amino acid
sequence of protein is called translation.

Translation starts from the point when rRNA molecule attached to one end of mRNA,
due to this attachment mRNA moves along ribosomes in increasing order of three
nucleotides. In the cytoplasm of a cell tRNA are present, which transfer specific amino
acid to ribosomes for protein synthesis. The tRNA attached with codon of mRNA with
the help of its anti-codon end.

As the mRNA moves along the ribosome adds an amino acid to growing polypeptide
chain. When ribosome moves to the next codon tRNA brings another amino acid. In this
way in each step an amino acid is added to the polypeptide chain. This process
continues until it meets the stop signal, which shows the end of polypeptide chain. It is
then separated from mRNA.

Steps of transcription
Step 1: Activation of amino acid

Amino acids must be activated for translation to occur. Activation ensures that the
correct amino acid will be recognized.

The amino acid is linked with ATP, liberating PPi and formed amino-AMP complex.

Amino acid + ATP ----------> Amino-AMP-enz + PPi

The amino group is enzymatically transferred to tRNA, liberating the enzyme Aminoacyl
tRNA Synthase and AMP.

Amino-AMP-enz + tRNA ----------> Amino acid + aminoacyl-tRNA synthase + AMP

Step 2: Formation of initiation complex

Formation of initiation complex require following three components

 A ribosome
 mRNA
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 tRNA carrying the first amino acid which is always methionine (Met)

These pieces form the initiation complex, the molecular setup needed to start making a
new protein.

Step 3: Elongation

Elongation is a key step of protein synthesis, during which the emerging polypeptide
chain extends by adding one amino acid.

During elongation the protein is synthesized one amino acid at a time on the 80S
ribosome. This process occurs in three major steps: binding of charged tRNA, peptide
bond formation, translocation of the growing peptide chain.

Step 4: Termination

When a stop codon appears the translation is terminated. There are no tRNA's that
recognize stop codons. Instead proteins called releasing factors, eRF, recognize the
stop codon. The releasing factors along with peptidyl transferases and GTP catalyze the
hydrolysis of the bond between the polypeptide chain and the tRNA. The protein and
tRNA disassociate from the site and the ribosome dissociates into the 40S and 60S
subunits releasing the mRNA.

GENETIC CODE
The information stored in the DNA is called genetic code or the sequence of bases
along the DNA strand serves as a code, known as genetic code. This information
translated into the amino acid sequence of proteins. The genetic code is like Morse
code.

Morse code: in Morse code letters are represented by combinations of long and short
signals of light or sounds.

The Genetic Code Uses Three Bases to Specify Each Amino Acid
The genetic information in the mRNA is composed of an alternating sequence of the
four nucleotide bases adenine (A), guanine (G), cytosine (C) and uracil (U) which
provides the unique code for specifying each of the 20 types of amino acids.

How 04 nucleotide bases are involved in coding for each type of amino acid? There are
only four kinds of bases and 20 kinds of amino acids. If only one base serve as a code
for an amino acid then we would have only 4 codes, if 2 bases serve as a code then we
would have (4)2 i.e. 16 possible codes but these codes were still not enough for 20
types of amino acids. In 1954 George Gamow a physicist from the University of
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Colorado (USA) pointed out that the minimum number of bases involved in amino acid
selection seemed to be three. This number gave a possibility of selection of (4)3 i.e. 64
combinations which are more than sufficient to specify 20 amino acids so, the code for
specifying an amino acid consist of 3-bases this is called triplet code. In 1961 Francis
Crick demonstrated that this hypothesis is correct. Most of the amino acids are specified
by more than one code. For example, 6 different triplet codons code for Arginine. (see
table)

It has been found that 61 out of 64 codes stand for amino acids and the rest three UAA,
UAG and UGA determines stop signals or termination point of the polypeptide chain
while the codons AUG and GUG not only code for Methionine and Valine but also
determines start signals. The DNA segment which is bounded by a start and stop
signals contains information for complete polypeptide chain is called cistron thus, a
cistron codes for protein, Sometimes, more than one adjacent cistrons code for a
polypeptide molecule and such a functionally related group of cistrons is called a gene.

Decoding
Decoding is the reverse of encoding or it is the transformation of information from
mRNA to amino acids sequence in polypeptide chain. In decoding ribosome exposes
the three-nucleotide sequence of mRNA, tRNA binds to codon of mRNA by its
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anticodon to determines the three-nucleotide sequence and carries the specific amino
acid for adding to polypeptide chain.

MUTATION
Mutations are sudden heritable changes in genes, chromosomes or genome of an
organism. Mutation is actually the change in the amount, content and organization of
genetic material. Genes or chromosomes transmit hereditary characters unchanged but
sometimes they undergo changes and alter the ability of cells to carry out biochemical
reactions and affect phenotype. Mutations are of two types i.e. chromosomal mutation
and gene mutation.

Chromosomal Aberrations
Chromosomal aberrations are concerned with the visible changes in the structure of
chromosomes or any structural change in the chromosome is called chromosomal
aberrations or chromosomal mutation.

Types of chromosomal aberrations

i- Deletion

Deletion is the loss of a part of a chromosome.

ii- Duplication

Duplication is a type of chromosomal aberration in which a segment of a chromosome becomes

part of another chromosome.

AB CDE FGH ------------> ABFGH ------------> Deletion

KLMNOP ------------> KLMCDENOP -------> Duplication

iii- Translation

In this type of chromosomal aberration exchange of segments between two non-homologous

chromosomes takes place.

ABCDEF ---------------> ABCNOP

KLMNOP --------------> KLMDEF

iv- Inversion

Inversion is a type of chromosomal aberration in which a reversal of segment of a chromosome

takes place.
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ABCDEFGH ----------------> ABFGHCCDE

Gene Mutation
Any change in number, structure and size of genes is called gene mutation.

Point Mutation
Mutation involving alteration in the sequence of one or few nucleotide is called point mutation.
Point mutation results from chemical or physical damage to DNA or from pairing error.

Transposition
The movement of gene from one place to another on chromosome is called transposition. When
gene moves to another location causes alteration or change in its expression. It may also
neighbor gene. When a segment of a chromosome changes their location or become part of
another chromosome is called chromosomal rearrangement. This chromosomal rearrangement
has a strong effect on the expression of the genetic message.

Mutagens
The chemicals produce and release by industries in environment and capable of DNA damage
called mutagens. OR

Any physical or chemical substance which can cause mutation in gene is called mutagens.

DNA Damage
Three major ways of DNA damage are as under

1) Ionizing Radiation

The radiation consisting of particles, X-rays, or gamma rays with sufficient energy to cause
ionization in the medium through which it passes is called ionizing radiation.

The X-rays and gamma rays can cause damage to DNA. The rays when reached to a cell
absorb by atoms and convey energy to electrons. When electron absorb the energy ejected out
from the atom. The ejected electron leaves an ionized atom with unpaired electron, each called
as free radical. The free radicals are highly reactive, reacting with DNA. The free radical breaks
the phosphodiester bond of the DNA. Mutational repair enzyme of the cell cannot repair this
damage. To repair the double strand break of DNA the two fragments should be fixed in the
position. The eukaryotes are able to position the two fragments by synaptonemal complex.
The synaptonemal complex is formed in the meiosis.

2) Ultraviolet radiation
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The ultraviolet rays are much lower in energy than X-rays. The ultra violet energy does not
convey enough energy that to eject electrons. Free radicals are not formed.

3) Chemical mutagens

Some chemicals can cause DNA damage. These chemicals are of three types.

DNA nucleotide like chemicals pair wrongly to DNA

Chemicals which remove amino acid from adenine and cytocine

Chemicals which add hydrocarbons to nucleotide

Examples of Gene Mutation

A) Sickle Cell Anemia

Sickle cell anemia is a heritable blood disease in which blood cells become sickle shaped in
comparison to the normal round shape. Due to sickle cell anemia individuals are unable to
transport oxygen to their tissues. When oxygen shorts the effective hemoglobin molecule
becomes insoluble and combines with one another, forming stiff rod like structure. This result in
the formation of sickle shaped red blood cells. This result in various abnormalities and may
ultimately result in death of individual.

The sickle cell hemoglobin differs from normal hemoglobin only in one amino acid in their chain.
In the normal Hb the chain has glutamic acid as the 6th amino acid where as in sickle cell
hemoglobin this is replaced by valine.

RBC’s with large proportion of defective molecule becomes sickle shaped and stiff. As a result
of this stiffness and irregular shaped structure of blood cells, blood cells move through blood
vessels with difficulty. In this way they may accumulate in the blood cells, forming clots.

This disease is caused by a mutation in a single gene which in heterozygous causes moderate
sickling when they are exposed to low level of oxygen and in homozygous condition causes
severe abnormality. The peoples suffering from sickle cell anemia have irregular illness and a
shortened life span.

B) Phenylketonuria

Phenylketonuria is a congential absence of phenylalanine hydroxylase (an enzyme that

converts phenylalanine into tyrosine). Due to this lack of enzyme the phenylalanine is not
broken down into tyrosine instead converted into a chemical that accumulate into blood and
seriously impair early neuronal development in infant. Phenylalanine interfere development of
brain cells and result in mental retardation and die before the age of 30 years. Phenylketonuria
is controlled by recessive allele of gene encoding the enzyme phenylalanine hydroxylase.