MUTATION

Microbial Genetics
STBP 2032
 Hypothesis : Mutation occurs RANDOMLY and is NOT INDUCED
by the exposure to antibiotics

This condition
109 cells/ml does NOTallow
Mixing between cells
in each tube
109 cells/ml

This condition
allows
mixing HIGHER fluctuation is observed ∴showed
that mutation occurs at different times or did not
occur at all in the different tubes
Figg 17.7 :
Rare tautomeric
shifts that occur
in the chemical
structure of the
four nitrogenous
bases of DNA
NOT STABLE,
can form
BUT RARE!
5- BROMOURACIL :
THYMINE
ANALOGUE
Nitrous Acid (HNO2):

Causes deaminations
Ethyl Methyl Sulfonate (EMS) :
Induced
mutation
b a chemical
by h i l
agent,
5-BU,
a T analogue
that pairs
with A

5-BU
causes
transition of
AT to GC
∴ Mutation
occurs
5-BU is a T analogue

5-BU undergoes TAUTOMERIC SHIFTS and therefore causes transition mutation

Where AT pairing changes to GC pairing and GC pairing changes to AT pairing
 NTG : alkylating
Mode of Action Agent That adds
(Mutation) By an alkyl group to
Nitrosoguanidine the N7 of guanine
(NTG) and causes a gap
in the base sequence
by deleting
Endonuclese enzyme cuts the DNA sugar- the purine ring
phosphate backbone at the 5’ end

(Gap filling)

Insertion of new
nucleotide
Acridine as a mutagen

Acridine does not

react with DNA,
DNA
But instead intercalates
Between dsDNA strands
u g DNA replication,
During ep cat o ,
Which causes Frame shift mutation :
Addition or deletion
of bases ATCC AGG GAT TAC

ATC CAG GGA TTA C

MUTATION CAUSED BY ULTRAVIOLET LIGHT RADIATION :
 MUTATION CAUSED BY ULTRAVIOLET LIGHT RADIATION :

Thymine dimers : causes DNA distortion, ∴

thi obstructs
this b t t transcription
t i ti
& induces mutation during
DNA replication

DNA repair In the presence of LIGHT :

(Photoreactivation enzyme)
Photoliase

Also requires one

light photon
LIGHT REPAIR : Mechanism of DNA repair due to damage by UV that occurs
in the presence Light is called PHOTOREACTIVATION
 MUTATION CAUSED BY ULTRAVIOLET LIGHT RADIATION :
DARK
REPAIR

Gap filling can cause errors in insertion of bases

UV can also cause breakage of DNA : thus
induces the loss of several base pairs

Breakage
Exonuclease
 Mutations Induced By Biological Mutagens :
TRANSPOSON
¾ Transposon : An element that can transfer from one location to another
new location in the genome
¾ Transposition : a process that transfers transposons which do not require
homology with the targeted site ∴ it is NOT site-specific
site specific

E. coli transposon,
p , Insertional Sequence
q (IS)
( ):

Transposon in a DNA
region

Insertion

Recipient DNA
(target DNA)
Different Kinds of Point Mutation
 ACRIDINE CAUSES FRAMESHIFT MUTATION

Gly Phe Gly

Misalignment

DNA
Repli-
cation

Insertion deletion
Gly Phe Trp Gly Leu
 REVERSION MUTATION

Sequence change
Frameshift Mutation

L th l
Lethal
(Its protein)
Silent
Reduced
Revert to the original genotype

Figure 10-15 Reversion by base deletion from an

acridine-induced base-addition
 REVERSION
(PHENOTYPE)

Amino acid remains

Mutant amino acid changed
g BUT reversion
changed & phenotype Amino
A i acid
id changed
h d tto positive
iti to wild type occurs, coz
reverted to wild type charge BUT genotype remained, mutation occurs affecting
Phenotype reverted to Wild type the other amino acid
Replica Plating : A method for screening/isolating
mutants (eg. Auxotrophic mutants ie in the studies of amino acid
biosynthesis
bi th i metabolism)
t b li )
Replica plating
 MUTATION IN A POPULATION

per 107 cells) 20

Mutation Rate = Mutation frequency
No of generation
No. of rresistant cells p

16 = a divided by b
Mutationn

a
Expressed in Units of :
mutants per 107
“mutants
ncy of M

12 cells per
generation” b
Frequen

8
(N
F

0 5 10 15 20 25
No. of generation/division
 However, if mutation does occur
at a rate of 2 x 10-8 (i.e.
(i e for each generation)
 Ames Test to determine the mutagenic
capabilities of a compound
¾The auxotrophic
p
strains were created
by point mutation,
therefore these
mutations can revert
back to WT by
Spontaneous or
induced Mutations

¾ These S.
S
typhimurium TA
strains can be used to
screen new
compounds
Agar plates used in Ames Test to
determine mutagenicity