Bacterial Genetics

Outline
Bacterial Genetics
Gene transfer
Genetic Recombination
Transposable elements
Significance of Bacterial genetics
 Contd
Genome total genetic material.
Bacterial genome Chromosome (circular &
haploid), circular plasmid, Transposon
Plasmids: fertility factors, resistance factors
circular, double stranded DNA,
replicate independently,
non essential function coding (additional)
Mutation
Contd
 DNA repair mechanism
4 mechanisms:
1. Excision repair: Nucleotide & Base
2. Recombination repair/ postreplication repair
retrieves missing information by genetic
recombination
3. SOS response
interruption of replication & correction of the
damage (error prone)
4. Error prone repair
Last resort before death; random sequence filling
 Genetic Expression
Transcription
is the transfer of DNA-bound protein synthesis instructions to mRNA.
is mediated in bacteria by RNA polymerase.
is initiated by the binding of sigma factor, a subunit of RNA polymerase, to the promoter
region of the DNA molecule.
occurs in a 5 PO4 to 3 OH direction.

Translation
occurs at the ribosome's.
is accomplished by the tRNA-mediated linkage of amino acids, in accordance
with the triplet-encoded mRNA transcript.
is the assembly of polypeptide chains from the mRNA transcript.
 Gene Regulation
Negative control
is inhibition of transcription by the binding of a repressor protein.
is exemplified by:
The lac operon
The trp operon
Positive control (catabolite activation)
is the initiation of transcription in response to the binding of an activator
protein
E.g. cyclic AMP receptor protein (CRP) enhances RNA polymerase
activity.
 Bacteriophage
RNA/DNA; single or double stranded encapsulated virus; needs host to replicate
Temperate phage; Virulent phage
 Genetic transfer
1.Conjugation
-Transfer of genes by cell to cell contact.
- Requires a cytoplasmic bridge between the cells.
- Both cells must have a proper genetic constitution to adhere to each other.
- Adherence requires the presence on the donor cell of a hair like projection
called sex pilus.
- occur in eubacteria,
- usually occur between members of the same or related species, but has
also been demonstrated to occur between prokaryotes and cells from
animals, plants, and fungi.

2. Transduction
Using a temperate phage vector without cell to cell contact.
i) Generalized Transduction:
Random fragment of bacterial DNA accidentally encapsulated in a phage
protein coat in place of the phage DNA.
The recipient incorporates the fragment by Recombination.
Generalized transducing particles should contain primarily bacterial DNA and
little or no phage DNA.
Generalized transducing particles are valuable in the genetic mapping of
bacterial chromosomes.
The closer two genes are within the bacterial chromosome, the more likely it is
that they will be co-transduced in the same fragment of DNA
ii) Specialized Transduction:
Certain bacterial genes close proximity to the phage are transduced.
The phage acquire these genes by a rare, abnormal excision from the bacterial
chromosome.
Unlike the above the phage contains both phage DNA & acquired bacterial
DNA as a single molecule.
The single molecule is incorporated by recombination
 3.Transformation
- Transfer of genes from one cell to another by means of naked
DNA.
- Discovery of transformation led to the identification of DNA as
genetic material.
- Competence ability of a cell to be transformed.
- Depends on physiologic state of the cell (ability of DNA crossing
cell membrane)
- As the free DNA enters the recipient, one the strand is
destroyed by Nucleases.
- The remaining single strand searches for a region of
homologous sequence and replaces one of the two resident
strands by a complex cut and paste process.
 GENETIC RECOMBINATION
An exogenote is conveyed into a recipient cell, its effect depends on what
happens after transfer. There are basically three possible fates.
1) The exogenote DNA may be degraded by a nuclease, in which case no
heritable change is brought about.
2) It may be stabilized by circularization and remain separate from the
endogenote
3) The third possible fate is recombination between exogenote and
endogenote, resulting in the formation of a partially hybrid chromosome
the two principal processes by which recombinant chromosomes are
formed following genetic transfer:
- homologous recombination and
- site specific recombination
 Homologous Recombination
Is a mechanism by which an exogenote can recombine with the bacterial
chromosome. This term reflects one of the two requirements for this
process:
(1) the exogenote must possess reasonably large regions of nucleotide
sequence identity or similarity to segments of the endogenote chromosome,
because extensive base pairing must occur between strands of the two
recombining molecules; and
(2) the recipient cell must possess the genetic ability to make a set of enzymes
that can bring about the covalent substitution of a segment of the exogenote
for the homologous region of the endogenote.
A protein known as RecA (recombination) controls the entire process.
Homologous recombination is responsible for integration of DNA
fragments transferred by generalized transduction, by plasmid-mediated
conjugation, and by natural transformation.
 Site-specific recombination
Are RecA independent, that rely on only limited DNA
sequence similarity at the sites of crossover
Are mediated by different sets of specialized enzymes
designed to catalyze recombination of only certain DNA
molecules.
The enzymes that bring about site-specific recombination
operate not on the basis of DNA homology but on recognition
of unique DNA sequences that form the borders of the
specific sites.
These enzymes are commonly encoded by genes on the
exogenote.
 Transposable elements
Genetic units capable of mediating their own
transfer
Dont exist as segments free of the genome
Relies on transposases (site specific
recombination enzymes)
3 major kinds: IS, Transposons & transposable
prophage
Contd
 Contd
Transposon:- has generally 3 genes
Transposase gene
Code for transposition
Resolvase gene
Antibiotic resistance gene
Transposition 2 types: Direct (replicative) & Non replicative
Insertion of transposon into functional gene loss of gene
function internal mutagenic agents.
Transposons
 The significance
In Genetic Engineering
In recent years, engineering
based on bacterial genetics has
transformed biology.
Many drugs can be formed by
using genetic engineering .
Proteins such as insulin that have
useful functions can be prepared
in large quantities from bacteria
that express cloned genes
 Significance cont..
Genetic transfer of antibiotic resistant genes
makes prescribed drugs ineffective especially for
mixed infections (alternate preparation of a new
drug)
E.g. antibiotics given for S. aureus , vancomycin, can not be
effective for individual infected by Enterococcus faecalis (UTI).