Mutations (L.

Muture – to change)
 Sudden inheritable discontinuous variations which appear in organisms due to
permanent change in their genotype

Types of Mutations
There are various schemes for classification of different kind of mutations. Depending
on

A. The Type of Cell Involved

Somatic mutations
 Mutations that are in the somatic tissues of the body.
 Mutations are not transmitted to progeny.
 The extent of the phenotypic effect depends upon whether the mutation is
dominant or recessive
 The extent of the phenotypic effect depends upon whether it occurs early or
late in development (early arising mutations have a greater effect).
Germinal mutations
 Mutations that are in the germ tissues of the body.
 Mutations may be transmitted to progeny
 Dominant mutations are seen in first generation after the mutation occurs
 If a female gamete containing an X-linked mutation is fertilized, the males will
show the mutant phenotype

B. based on cause of mutation
2 kinds of mutations- chromosomal mutations & gene mutations
1. Chromosomal Mutation and Types
 The changes in the chromosome parts and chromosome sets are called
chromosome aberrations or chromosome mutations.
 types: Changes in chromosome numbers & Change in chromosome structure

Changes in chromosome numbers – 2 types: Polyploidy & aneuploidy

Polyploidy
 Having more than 2 set of chromosomes in diploid organisms
 Failure of chromosome separation during anaphase of meiosis – would resulted in
polyploidy - Triploid (3n) & tetraploid (4n)
Aneuploidy
 Having additional or fewer chromosomes
 Hypoploid – loss of Chromosome (n-1)
 Hyperploid – addition of chromosome (n+1)

Change in chromosome structure/Chromosomal aberrations

 Aberrations/Structural change within same chromosome – changes can be
classified as:
o Deletion – loss of middle segment of a chromosome
o Inversion – part of the chromosome inverted
o Translocation - Movement of a segment into a non-homologous
chromosome
o Duplication – one segment of chromosome got duplicated
2. Gene Mutations
 Mutations resulted in changes in number, sequence and type of nucleotide
 Developed due to error in DNA

I. based on nature or mode of mutations

two types of mutations have been recognised
1. Point mutation - alterations occur in a single nucleotide or nucleotide pair
The point mutations may occur due to following types of nucleotide change
Deletion mutations
 caused due to loss or deletion of some portion (single nucleotide pair)
 Deletion mutations have been frequently reported in some
bacteriophages
Insertion or addition mutation
 occur due to addition of one or more extra nucleotides
 The insertion - induced by certain chemical substances
Substitution mutation
 A point mutation in which a nucleotide is replaced by another nucleotide
 That altered nucleotide may affects the reading frame
 Substitution mutations – occurs in two ways
o Transition - When a purine (e.g., adenine) base is substituted by
another purine base (e.g., guanine) or a pyrimidine (e.g., thymine) is
substituted by another pyrimidine base, (e.g., cytosine)
o Transversion – Substitution or replacement of a purine with a
pyrimidine or vice versa
2. Multiple mutations or gross mutations
 When changes involving more than one nucleotide pair, or entire gene - gross
mutations. The gross mutations occur due to rearrangements of genes within the
genome. It may be:
o rearrangement of segment of genes
o rearrangement of number of genes

II. Based on effects of mutations

3 kinds of mutations
1. Non-sense mutations - mutations convert a codon that originally specified an amino
acid into one of the three nonsense codons or stop codons UAA, UAG & UGA.
2. Missense mutations - mutation that change a codon so that it specifies a different
amino acid. Frameshift mutations, by additions or deletions of nucleotides, cause an
alteration in the reading frame of codons
3. Silent mutations - mutations are those that do not alter the amino acid sequence of
the polypeptide even though the nucleotide sequence has changed.
C. Based on origin of mutations
1. Spontaneous mutations
 The spontaneous mutations occur suddenly in the nature and their origin is
unknown. They are also called “background mutation”
 Rare one (1 in 2000 to several million Nucleotides)
 Most of the mutations occur due to error in DNA proof reading mechanism
after replication

2. Induced mutations
 Besides naturally occurring spontaneous mutations, the mutations can be induced
artificially in the living organisms by exposing them to abnormal environment
such as radiation, certain physical conditions (i.e., temperature) and chemicals.
 Muller (1927) – used X rays to induce mutations in Drosophila
 An agents that induce the mutation in the gene or genome generally known as
“Mutagens” – based on nature of mutagens, it can be classified into 3 types: (i)
Physical mutagens (ii) Chemical mutagens & (iii) Biological mutagens
 Physical mutagens – 2 physical agents are responsible for mutations
 Temperature – increase in temperature, increases the rate of
mutations; rise in temperature generally break the hydrogen bond
between bases and increases the possibility of mispairing of bases
 Radiations – radiation such as X-rays, gamma, beta rays & UV rays
are also induce mutations when exposed to them
UV rays – Non-ionising radiation – responsible for thymine dimer
formation
X-rays – Ionising radiation – deamination or dehydroxylate the
nuclotides – mis-match pairing of nucleotides
 Chemical mutagens – several chemicals are induce mutations – the
following group of chemicals are responsible for mutations
 Deaminating agents – removal of amino group from bases especially
from A,G & C. Nitrous acid [HNO2] – conversion of A to G
(Transition); and change the base pairing from A=T to G=C
 Alkylating agents – donate alkyl groups to reactive groups of bases
Resulted in methylation & ethylation of nitrogenous bases; all
kinds of mutations including transition, transversion, & frameshift
Ex. Nitrogen mustard, ethyl methane sulfonate (EMS) & methyl
methane sulfonate (MMS)
 Base analogs – certain chemicals structurally similar to nucleotides
and wrongly incorporated into DNA instead of normal bases.
 Incorporation of such analogs – alters the pairing of nucleotides
Alteration of bases later alters the transcription & translational
products.
 Intercalating agents – certain positively charged molecules
intercalate and stock between nucleotides in the DNA and
facilitates the error during replication
Ex. Acridine dyes & EtBr.
 Biological agents – viral genes got integrated into host genome during
infection and alter the arrangement of genes.

Ames Test
Ames test it is a biological assay to assess the mutagenic potential of chemical
compounds. It utilizes bacteria to test whether a given chemical can cause mutations in
the DNA of the test organism. The test was developed by Bruce N. Ames in 1970s to
determine if a chemical at hand is a mutagen.

Principle
1. Ames test uses several strains of bacteria such as Salmonella and E.coli that
carry a particular mutation.
2. Point mutations are made in the histidine (Salmonella typhimurium) or the
tryptophan (Escherichia coli) operon, rendering the bacteria incapable of
producing the corresponding amino acid.
3. These mutations result in his- or trp- organisms that cannot grow unless histidine
or tryptophan is supplied.
4. But culturing His- Salmonella is in a media containing certain chemicals, causes
mutation in histidine encoding gene, such that they regain the ability to
synthesize histidine (His+). This is to say that when a mutagenic event occurs,
base substitutions or frameshifts within the gene can cause a reversion to
normal. This is the reverse mutation.
5. These reverted bacteria will then grow in histidine- or tryptophan-deficient
media, respectively.

A sample’s mutagenic potential is assessed by exposing amino acid-requiring organisms

to varying concentrations of chemical and selecting for the reversion event. Media
lacking the specific amino acid are used for this selection which allow only those cells
that have undergone the reversion to histidine / tryptophan prototrophy to survive and
grow. If the test sample causes this reversion, it is a mutagen.
Merits
 Simple, rapid and robust bacterial assay.
 Ease and low cost of the test make it invaluable for screening substances in our
environment for possible carcinogenicity.
 Ames test can detects suitable mutants in large population of bacteria with high
sensitivity.
Limitations
 Some substances that cause cancer in laboratory animals (dioxin, for example)
do not give a positive Ames test (and vice-versa)
 Ames assay consists of Salmonella typhimurium strains and so it is not a perfect
model for human.