Bacterial Genetics

 Definitions
Genetics: is the science of heredity and
variation.
Heredity: implies the transmission or
inheritance of constant similar features
between generations. (Like father like
son)
Variation: refers to the fact that similar
organisms can be differentiated from
each other parameters for
differentiation may be size, coloration,
morphology, behaviour, their physiology
and biochemistry.
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 Gene: is a hereditary unit,

that occupies a specific location on a
chromosome,
determines a particular characteristic
in an organism by
directing the formation of a specific
protein, and
is capable of replicating itself at each
cell division.

 A Gene can also be defined as a unit

or chemical entity that gives rise to,
or controls a trait under a given set
of environmental conditions.
Trait: is any feature about an
organism that can be observed.

 Phenotype: The phenotype of an

individual organism is either its total
physical appearance and constitution or
a specific manifestation of a trait, such
as size, eye color, or behavior that
varies between individuals. Phenotype
is determined to some extent by
genotype.
Genotype: is the genetic constitution
of the organism.
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 Interaction between genotype and phenotype in a

simple equation:
genotype + environment phenotype
Eg
Diabetic genotype

Diabetic genotype

environment

diabetic phenotype

Normal diet
environment

normal /near normal phenotype

Special diet /
Antidiabetic drugs

= a phenotype associated with a given genotype

may vary under different environmental conditions.

Bacterial genetics-:

the study of how the

properties of bacteria can be
explained by the specific
genes that the bacteria
contain.

 medically, we are concerned

with genes that determine basic cellular
processes,

central metabolism
ability to cause disease in man, and to
resist the action of antibiotics.
contributions to medicine:
rational design of new
antimicrobial compounds, &
the development of highly
effective vaccines.
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variation

bacteria like other organisms breed true and

maintain their characters constant from
generation to generation,
a small proportion of their progeny show
variations of particular characters.
The phenotypes (observed character) of a
pure growth are liable to two kinds of
variation:
Temporal variations due to changes in
environment.
(These changes are usually
non-inheritable).
Permanent variations (mutation) which are
inheritable.

Mutation: Spontaneous and

induced

In a pure culture, one can find

thousands of mutants that will have
different genotypes as compared to
the parent cell.
This results from spontaneous
mutations that occur in the culture.
cause of spontaneous mutation is
not known.
mutation can be induced in a culture
by exposing it to X ray, UV light or
other forms of radiation.
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 Eg.penicillin production, mutants of

Penicillium chrysogenum with better yields.
Certain chemical agents (mutagens) can
cause induced mutation (eg antibiotic
resistance development in chemotherapy).
Mutation induced by such agents do not
become immediately evident but only after
several generations when the changed part
of the genetic material has become
segregated in the newly formed cells and
exhibit their controlling influence on their
properties and behaviour.
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DNA & Genetic information

Nucleic acids bear genetic
information of organisms (DNA &
sometimes RNA)
Implies that DNA codes for genetic
composition and also
controls the activity/ties of
organisms.
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Structure of DNA
Purine bases: adenine, guanine
Pyrimidine bases: thymine, cytosine
Deoxyribose (sugar), PO4
Diagram illustrating a nucleotide, the building blocks of DNA

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 Genetic information is stored in DNA

by different sequential arrangement
of the purine and pyrimidine
nucleotides.
there are thousands of these
compounds in each molecule of DNA
this makes it possible to store a
great deal of information in one DNA
molecule.
One sequential arrangement may
control one reaction of the cell.
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 the DNA serves as a template for synthesis

of more DNA (which will normally have
identical structure)
any error in the replication of the paired
purine pyrimidine sequence changes the
metabolic reaction being controlled by the
original DNA.
this is thought to be the cause of mutation.
In bacteria there is only one large molecule
of DNA (unlike in eukariotic cells = double
helix)
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An experiment to illustrate that DNA

bears Genetic Information
Experiments by Frederick Griffith
(1928) found there were two
different types of the bacterium
Streptococcus pneumoniae:
An "S" or SMOOTH coat strain, which
is lethal to mice.

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An "R" or rough strain, which will not

hurt the mouse.

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Griffith found that he could

heat inactivate the smooth
strain.

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However, if he were to take a mixture of the heat-inactivated

S strain,
mixed with the R strain, the bacteria would die. Thus there
was some
Material in the heat-killed S strain that was responsible for
"transforming" the R strain into a lethal form.

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 Fred Griffith (and a lab co-worker) was

killed in their laboratory in 1940 from a
German bomb. However, their work
continued on in the U.S. in 1944, by
Oswald Avery, C.M. MacLeod, and M.
McCarty carefully demonstrated that
extracts from the dead cells were
responsible for the transformation.
The chemical substance in the extract was
shown to be deoxyribonucleic acid (DNA)
The mutagen could be inactivated by the
specific enzyme - deoxyribonuclease.
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Averys expt
Component of the extract causing the
transformation:
Proteins or DNA
R. strain + S. strain extract + Protease =
transformation occurred
R. strain + S. strain extract + DNAase =
No transformation
conclusion: DNA (and not proteins) is the
heredity molecule.
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Alfred Hershy & Martha Chase (1952)

Another expt to prove DNA as the source
of heredity information:
bacteriophage
Use 2 radioactive labels to track the
movement of
Protein (isotope 35S) &
DNA (isotope 32P) separately.

Expt: 35S-phage (labeled protein) added

to growing culture of E coli.
Obs: 35S-label remains outside of the
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bacteria cells

 Expt: 32P-phage (labeled DNA) added to

growing culture of E coli.
Obs: 32P-label enters interior of the
bacteria cells
Conclusion: DNA was the material that
enters the cells & therefore must be the
substance that carried the heredity
information.
Progeny bacteriophages: contained
and not 35S.
Shows that DNA passes from one
generation to the next.

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Plasmids
Plasmids are small extra chromosomal
pieces of genetic materials that can
replicate autonomously and maintain
themselves in the cytoplasm for
generations.
Plasmids can be transferred from one
bacteria to another (even of different
spp).
They are usually non essential to the cell.
Plasmids are commonly found in a
number of gram negative bacilli
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especially enterobacilli

 Plasmids can contain genetic information

that determines the ability of bacteria to
mate and ( also to act as the donor during
mating).
that codes for resistance to antibiotics &
other chemicals.
that code for the degradation of various
complex organic cpds.
for toxin production (in pathogenic bacteria).
One bacterial cell can contain more that
one plasmid
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Gene
transfer
The
methods by which genes can be
transferred in bacteria include
Phage-mediated transduction
The temperate phage before lysis picks up some
of the genetic material of the lysogenic
bacterium.
Later when the bacterial cell lyses it releases
phages some of which contain the genetic
material picked from the cell.
When such phages re-infect other bacterial cells,
these take on new characteristics owing to the
integration of the genetic material of the phage
into the recipients genetic make up.
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 Phage conversion
When a susceptible bacterial culture is
infected with a phage every cell that
becomes lysogenic acquires new
properties imparted by the infecting
phage.
They are retained by the progeny for as
long as these are parasitized by the phage
in the prophage state and the prophage
reproduces in synchrony with the bacterial
chromosome.
If the prophage is lost, the lysogenic stage
ends and the acquired property (ties) are
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lost.

 Conjugation
requires the presence of a genetic element,
the fertility or F factor.
The cell containing F factor is called the F+.
The female strains (F-) do not contain the F
factor.
a bridge forms between specific pilli (sex pilli)
on the F+ strain and receptor site on the F-.
genetic material passes only from the F+ to
the F- bacterium.
The genetic material of F+ may becomes
integrated into the chromosome of F-.
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 During the transfer across the bridge it is

possible for the bridge to break at any time.

The fertility factor is infectious.

In F+ males the factor is in the

cytoplasm and during conjugation can
infect F- bacterium.
Another type of male bacterium is
referred to as Hfr (high frequency of
recombination),
in this the F factor is integrated into the
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bacterial genetic material.

 Hfr is not infectious,

transfer of chromosomal material
occurs more frequently than in F+
males
few of the recipients of Hfr become F+
themselves.

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 Episome: a term postulated to explain the

mechanisms for hereditary material transfer in
transduction and conjugation.
Episomes are genetic structures containing DNA
that may be present or absent in bacteria.
They can exist in two alternative states:
a) free in the cytoplasm and multiply
independently of cell division or
b) integrate with genetic material and multiply
only when the genetic material replicates.
An integrated episome may become
cytoplasmic and a cytoplasmic episome may
become integrated.
Thus F factor may be present and when present
be free (F+) in cytoplasm or bound as in Hfr.
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Summary, genetic transfer

Genetic material can be transferred
between bacteria by several means,
most often by:
Conjugation
Transformation, and
Transduction
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 Conjugation
Is mediated by a particular kind of
circular DNA called F factor (a plasmid),
which replicates independently of the
chromosome.
When two cells are in close proximity to
each other, a hollow bridge-like structure,
forms between two cells.
This allows a copy of the plasmid, as it is
duplicated, to be transferred from one
bacterium to the other.
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 Transformation
During this process, genes are
transferred from one bacterium to
another as naked DNA.
When cells die and break apart, DNA can
be released into the surrounding
environment. Other bacteria in close
proximity can scavenge this free-floating
DNA, and incorporate it into their own
DNA. This DNA may contain
advantageous genes,
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 Transduction
In this process, bacterial DNA is
transferred from one bacterium to
another inside a phage that infects
bacteria.
When a phage infects a bacterium, it
essentially takes over the bacteria's
genetic processes to produce more
phages.

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 In this process of phage infection,

bacterial DNA may inadvertently be
incorporated into the new phage DNA.
Upon bacterial death and lysis, these
new phages go on to infect other
bacteria.
A phage brings along genes from the
previously infected bacterium.
The imported genes from the phage
incorporates into the genome of the
new bacterium.
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Thank you.

GOOD LUCK!!!
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