Microbial Genetics
BP 605 T. Pharmaceutical Biotechnology (Theory)
PRESENTED BY – NANDANI THAKUR
Overview
Introduction to Microbial genes
Mutations and types
Gene transfer
Microbial Genetics
DEFINATION: Microbial genetics is the study of the mechanisms of heritable information in
microorganisms, including bacteria, archaea, viruses and some protozoa and fungi
Strain or clone: A clone is a population of cells that are genetically ideal pure culture.
Genome : All the genes present in a cell.
Phenotype: Collection of characteristics that are observable.
Genotype: Specific set of genes it possess.
Gene: A gene is a nucleotide sequence that code for a polypeptide, tRNA or rRNA. Most bacterial
genes have at least four major parts promoters, leaders, coding regions and terminator.
Genetic recombination: Two separate genomes are brought together in one unit.
Gene Expression
The expression of a gene into a protein occurs by:
1) Transcription of a gene into RNA
• produces an RNA copy of the coding region of a gene
• the RNA transcript may be the actual gene product (rRNA, tRNA) or be translated into a
polypeptide gene product (mRNA)
2) Translation of mRNA transcript into polypeptide
• accomplished by ribosomes with the help of tRNA
Overview of Gene expression
DNA Replication
Features of DNA Replication
Both strands serve as a template:
• synthesis is always 5’-3’
• leading strand synthesis is continuous,
lagging strand synthesis is discontinuous
Each new DNA fragment requires an RNA primer:
• DNA synthesis cannot begin without a primer to add
to
Some important enzymes:
DNA Polymerase (synthesizes new DNA)
Primase (makes RNA primers)
DNA Ligase (“stitches” fragments together)
Genetic recombination or Gene Transfer in Bacteria
In prokaryotes, genetic recombination occurs because fragments
of homologous DNA from a donor chromosome are transferred to
a recipient cell by any of the three following processes.
• Transformation – Transfer of bacterial genes involving free DNA.
• Transduction – Transfer of host genes from one cell to another medicated by a virus.
• Conjugation – Transfer or genes from one cell to another involving cell to cell contact
and a plasmid.
Transformation
• A cell that is able to take up a molecule of DNA and be transferred is called
competent cell.
• Bacteria differ in the form in which DNA is taken up. In Gram negative
bacteria (eg. Haemophilus) only DS DNA is taken up into the cell,
however only SS – DNA segment is incorporated into the genome.
• In Gram positive bacteria (Streptococcus sp. and Bacillus) only SS – DNA is
taken up.
Transformation
Under the right conditions, bacteria can “take
in” external DNA fragments (or plasmids) by
transformation.
• DNA binding proteins
transfer
external DNA across cell envelope
• homologous recombination can then
occur
• bacterial cells capable of transformation are
referred to as competent
Transduction
• Not all phages can transduce and not all bacteria are transducible.
• In generalized transduction host DNA derived from any portion of host
genome becomes a part of the DNA of the mature virus particle in place of the
virus genome, which gets integrated into another cell upon entry.
• In specialized transduction, when a lysogenized cell reverts to lytic cycle, a
part of host DNA is exchanged for phage DNA, which replicates and forms
phage, which when trasnduced, the new gene gets into another cell.
Bacterial Transduction
Conjugation
• Conjugation or mating – involves the transfer of DNA from a donor to a
recipient by cell to cell contact through the F (Fertility) pilus, followed by
recombination within the recipient bacterial cell.
• Pili are involved in attachment processes.
• F pili specifically join mating bacteria. When an F pilus joins with the mate,
there is a change in plasma membrane permeability so that DNA can
move from one cell to another.
• Bacteria that produce F pili are donors and are designated F+ strains.
Conjugation
During mating, a single strand of donor DNA is replicated, and this copy is
transferred to the recipient where the complimentary strand is synthesized.
Bacteria are designated Hfr (high frequency recombinant) if the F
plasmid DNA is incorporated into the bacterial chromosome.
Bacteria lacking F pili are recipient strains and are designated F –
strains. When F+ cell mates with F– cell, the F plasmid DNA is copied and
transferred from donor to the recipient. This results in F+ strains.
The F plasmid confers the genetic information for acting as a donor strain.
Bacterial Conjugation
Requires an F factor plasmid
• has all “conjugation genes”
• directs formation of a sex pilus
• single DNA strand produced by DNA replication is
transferred to F- cell through the sex pilus,
recipient produces 2nd strand
Bacterial conjugation
Plasmid
• Some bacterial cells contain one or more small circular macromolecules of
DNA that store additional specialized information. These are called
plasmids (extra chromosomal DNA).
• Plasmids contain only 1 – 5% as much DNA as in the bacterial chromosome
(roughly about 20 genes) which supplement the essential
genetic information contained in the bacterial chromosome.
Plasmid
• Genetic information contained in plasmids can be important, in
establishing characters such as resistance to antibiotics and tolerance
to heavy metals.
• Thus the gene products of plasmids may permit the survival of bacteria
under conditions that are normally unfavourable for growth and survival.
• Plasmids can be transferred from one bacterial cell to another, sometimes
even from one bacterial species to another.
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