Bacterial Genetics

BACTERIAL GENETICS
I MSc Botany Based on Prescott
Dr Giby Kuriakose
Key Words
Genetics Bacteriophage
Bacterial genetics Lysogenic cycle
Mutation & its types Mechanisms of gene
Point mutation transfer
Frameshift mutation Transformation
Lethal mutation Transduction
Suppressor mutation Lysogenic conversion
Missense & nonsense Conjugation
mutation Transposition (Jumping
Genes)
Bacterial Genetics
Genetics is the study of heredity and variation.
The unit of heredity is gene, which is a segment

of DNA specifying for a particular polypeptide.
Introns - non coding sequences on a gene.
Exons - coding sequences on a gene translated into

gene products.
Bacterial genetics is used as a model to understand

DNA replication, genetic characters, their changes &
transfer to next generations.
Nucleic Acids
DNA ( deoxy ribonucleic acid ) : stores
information for protein synthesis.
RNA ( ribonucleic acid ) : transcription &

translation of information for protein synthesis.
Central Dogma : DNA RNA Protein

Structure Of DNA
Proposed by Watson & Crick.
Double helix model.
Composed of 2 chains of polypeptides, each
chain has a backbone of deoxyribose sugar and
phosphate residues arranged alternately.
4 nitrogenous bases: Adenine (A) Purine
Guanine (G)
Thymine(T) Pyrimidine
Cytosine (C)
Double helical
structure of DNA by
Watson & Crick
Structure Of RNA
Structurally similar to DNA, except for 2
major differences:
ribose sugar
uracil in place of thymine.
3 types of RNA
m RNA (messenger RNA)
t RNA ( transfer RNA )
r RNA ( ribosomal RNA )
Genetic Information In Bacteria
Chromosome Carries properties like virulence,
pathogenicity & resistance
Plasmid Extrachromosomal genetic

material in the cytoplasm
Replicate independently
Bacteriophage Virus infecting bacteria

PLASMIDS
Circular DNA molecules
Important vectors in genetic engineering
EPISOME
Plasmid DNA integrated with chromosomal DNA.
Types of plasmids
R plasmid (drug resistance): RTF* + r determinant
F plasmid (maleness )
* Resistance Transfer Factor

Genotypic & Phenotypic Variations
Genotype – genetic constitution of a cell that is
transmitted to its progeny
Phenotype – physical expression of the genotype

in a given environment
Variations
1. Phenotypic variations –
influenced by the environment
temporary & not heritable
2. Genotypic variations –
Not influenced by the environment
Stable & heritable
Mechanisms Of Genetic Variations
Mutation
Transfer or exchange of genetic material

1. Transformation
2. Transduction
3. Conjugation
4. Lysogenic conversion
5. Transposition
Mutation
Random, undirected heritable variation
Caused by a change in the nucleotide base

sequence of the DNA
Types of mutation:
1. Point mutation
2. Frame shift mutation
3. Lethal mutation
4. Suppressor mutation
Mutagens - Agents which can induce mutation e.g.

UV rays, 5 bromouracil, alkylating agents, etc.
1. Point Mutation
Cause - due to addition, deletion or
substitution of one or more bases.
Types -
Transition : a purine base is replaced by a
purine base or a pyrimidine base is replaced
by another pyrimidine base.
Most common type.
Transversion : substitution of a purine base by

a pyrimidine base & vice versa
1. Point Mutation
Results of mutation -
Missense mutation – triplet code is
altered so that a different amino
acid is present at a particular
position in the protein.
Nonsense mutation – converts a

codon that specifies an amino acid
into a termination codon.
2. Frame Shift Mutation
Cause - Deletion or
insertion of a base -
changes all of the
codons downstream
from the change
3. Lethal Mutation
Mutation which resulting involve vital functions

in the death of the organism – nonviable
mutation.
A conditional lethal mutant may be able to live under

certain conditions – permissive conditions.
Commonest type of conditional mutant is the

temperature sensitive (ts) mutant which is able to live
at the permissive temperature of 35°C but not at the
restrictive temp (39°C).
Suppressor Mutation
Reversal of a mutant phenotype by another mutation at a

position on the DNA, distinct from that of the original
mutation.
Lederberg & Tatum (1946) Experiment demonstrating recombination in
E. coli. Recombination of 2 complimentary auxotrophs gives rise to a
strain that can synthesize all nutrients.
Bernard Davis experiment demonstrated that physical contact is
required for bacterial recombination.
Conjugation-transfer of the sex factor F:
1. William Hayes (1953) demonstrated that genetic

exchange in E. coli occurs only in one direction.
2. Genetic transfer is mediated by sex factor F.
3. Donor is F+ and recipient is F-.
4. F is a self-replicating, circular DNA plasmid (1/40

the size of the main chromosome).
5. F plasmid contains an origin sequence (O), which

initiates DNA transfer. It also contains genes for
hair-like cell surface (F-pili or sex-pili), which aid in
contact between cells..
F factor and Conjugation
F (fertility) factor is a conjugative plasmid transferred

from cell to cell by conjugation
F factor is an episome = genetic element that can insert

into chromosome or replicate as circular plasmid
The F plasmid is a low-copy-number plasmid ~100 kb in

length, and is present in 1–2 copies per cell
It replicates once per cell cycle and segregates to both

daughter cells in cell division
21
F factor and Conjugation
• Conjugation is a process in which DNA is transferred

from bacterial donor, F+ cell to a recipient, F- cell by
direct contact.
• The transfer is mediated by a tube-like structure called

a pilus, formed between the cells, through which the
plasmid DNA passes.
• Once in contact, conjugation, DNA transfer is

unidirectional. The lagging strand template peels
away… and is transferred to the recipient.
• The leading strand template is replicated in the donor

while the lagging strand template is replicated in the
recipient… so that both cells wind up with the plasmid.
22
1. No conjugation can occur between cells of the same
mating type.
2. Conjugation begins when the F plasmid is nicked at

the origin, and a single strand is transferred using
the rolling circle mechanism.
3. When transfer is complete, both cells are F+ double-

stranded
Transfer of the F factor
25
Conjugation of high-frequency recombinant strains:
1. No chromosomal DNA is transferred by standard sex

factor F.
2. Transfer of chromosome DNA is facilitated by special

strains of F+ integrated into the bacteria chromosome
by crossing over.
3. Hfr strains = high frequency recombination strains.
4. Discovered by William Hayes and Luca Cavalli-Sforza.
5. Hfr strains replicate F factor as part of their main

chromosome.
1. Conjugation in Hfr strains begins when F+ is nicked
at the origin, and F+ and bacteria chromosomal DNA
are transferred using the rolling circle mechanism.
2. Complete F+ sequence (or complete chromosomal

DNA) is rarely transferred (1/10,000) because
bacteria separate randomly before DNA synthesis
completes.
3. Recombinants are produced by crossover of the

recipient chromosome and donor DNA containing F+.
Transfer of the Hfr F+ factor
Excision of the F+ factor also
occurs spontaneously at low
frequency.
1. Begin with Hfr cell containing

F+.
2. Small section of host

chromosome also may be
excised, creating an F’ plasmid.
3. F’ plasmid is named for the

gene it carries, e.g., F’ (lac)
Transformation (Griffith, 1928)
Transfer of genetic information by free DNA. i.e. by
direct uptake of donor DNA by the recipient DNA.
Live noncapsulated (R) pneumococci + heat killed

capsulated (S) pneumococci
Injected into mice
Death of mice
Live capsulated pneumococcus isolated from the

blood of mice.
Transduction
Transfer of a portion of the DNA from one bacterium to
another by a bacteriophage.
Packaging error within the infected bacteria during the

assembly of progeny phages – presence of a segment of
host DNA along with the phage nucleic acid in the core of
phage
Infection of another bacterium
Transfer of host bacterial DNA to the new bacterium
Acquisition of new characteristics coded by the donor DNA.

Transformation
1. Unidirectional transfer of extracellular DNA into

cells, resulting in a phenotypic change in the
recipient.
2. First discovered by Frederick Griffith (1928).
3. DNA from a donor bacteria is extracted and

purified, broken into fragments, and added to a
recipient strain.
4. Donor and recipient have different phenotypes and

genotypes.
5. If recombination occurs, new recombinant

phenotypes appear.
Transformation:
1. Bacteria vary in their ability to take up DNA.
2. Bacteria such as Bacillus subtilis take up DNA

naturally.
3. Other strains are engineered (i.e., competent

cells).
4. Competent cells are electroporated or treated

chemically to induce E. coli to take up
extracellular DNA.
Transformation of Bacillus subtilus
Heteroduplex DNA
Transduction
1. Bacteriophages (bacterial viruses) transfer genes to
bacteria (e.g., T2, T4, T5, T6, T7, and λ).
1. Generalized transduction transfers any gene.
1. Specialized transduction transfers specific genes.
2. Phages typically carry small amounts of DNA, ~1% of

the host chromosome.
3. Viral DNA undergoes recombination with homologous

host chromosome DNA.
4. Most widely used mechanism of gene transfer among

prokaryotes
Fig. 15.12
Life cycle of phage λ

Fig. 15.13
Generalized transduction
of E. coli by phage P1
Lysogenic Conversion
Phage DNA itself is the new genetic element.
Bacteriophages – 2 Types of life cycle
Lytic or virulent cycle – progeny viruses build

up inside host bacterium, which rupture to
release them.
Temperate or nonlytic or lysogenic cycle –

host bacterium is unharmed.
Lysogeny
Lysogenic bacteria
Prophage behaves as an additional segment of

bacterial chromosome, coding for new
characteristics. This process by which prophage
confers genetic information to a bacterium is
called Lysogenic conversion .
Conjugation
First described by Lederburg & Tatum in 1946 in a strain of E.coli
called K12.
A donor or male bacterium passes DNA directly to a recipient or

female bacterium by a conjugation tube (sex pili). The female
bacterium attains donor status & in turn can conjugate with other
female cells.
Maleness is determined by the presence of a plasmid which

codes for sex pili.
The plasmid is called the sex factor or fertility factor (F factor)
R (resistance) factor can also be transferred by conjugation

Process of Conjugation
Transposon (Jumping Genes, Barbara McClintock)
DNA segment that can move Plasmid

between chromosome & plasmids
Chromosome
Insertion of transposon into a functional

gene would destroy the function of the
gene (internal mutagenic agents)
Transposon
Transposons are not self replicative, they depend on chromosomal or

plasmid DNA for replication

Bacterial Genetics

Uploaded by

Copyright:

Available Formats

Bacterial Genetics

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Bacterial Genetics

Uploaded by

Copyright:

Available Formats

BACTERIAL GENETICS

I MSc Botany Based on Prescott

The unit of heredity is gene, which is a segment

Introns - non coding sequences on a gene.

Exons - coding sequences on a gene translated into

Bacterial genetics is used as a model to understand

RNA ( ribonucleic acid ) : transcription &

Central Dogma : DNA RNA Protein

Plasmid Extrachromosomal genetic

Bacteriophage Virus infecting bacteria

Important vectors in genetic engineering

* Resistance Transfer Factor

Phenotype – physical expression of the genotype

Transfer or exchange of genetic material

Caused by a change in the nucleotide base

Mutagens - Agents which can induce mutation e.g.

Transversion : substitution of a purine base by

Nonsense mutation – converts a

Mutation which resulting involve vital functions

A conditional lethal mutant may be able to live under

Commonest type of conditional mutant is the

Reversal of a mutant phenotype by another mutation at a

1. William Hayes (1953) demonstrated that genetic

2. Genetic transfer is mediated by sex factor F.

3. Donor is F+ and recipient is F-.

4. F is a self-replicating, circular DNA plasmid (1/40

5. F plasmid contains an origin sequence (O), which

F (fertility) factor is a conjugative plasmid transferred

F factor is an episome = genetic element that can insert

The F plasmid is a low-copy-number plasmid ~100 kb in

It replicates once per cell cycle and segregates to both

• Conjugation is a process in which DNA is transferred

• The transfer is mediated by a tube-like structure called

• Once in contact, conjugation, DNA transfer is

• The leading strand template is replicated in the donor

2. Conjugation begins when the F plasmid is nicked at

3. When transfer is complete, both cells are F+ double-

1. No chromosomal DNA is transferred by standard sex

2. Transfer of chromosome DNA is facilitated by special

3. Hfr strains = high frequency recombination strains.

4. Discovered by William Hayes and Luca Cavalli-Sforza.

5. Hfr strains replicate F factor as part of their main

2. Complete F+ sequence (or complete chromosomal

3. Recombinants are produced by crossover of the

1. Begin with Hfr cell containing

2. Small section of host

3. F’ plasmid is named for the

Live noncapsulated (R) pneumococci + heat killed

Injected into mice

Live capsulated pneumococcus isolated from the

Packaging error within the infected bacteria during the

Infection of another bacterium

Transfer of host bacterial DNA to the new bacterium

Acquisition of new characteristics coded by the donor DNA.

1. Unidirectional transfer of extracellular DNA into

2. First discovered by Frederick Griffith (1928).

3. DNA from a donor bacteria is extracted and

4. Donor and recipient have different phenotypes and

5. If recombination occurs, new recombinant