Bacterial Genetics

BY

DR YUGUDA S.
@
DEPARTMENT OF MEDICAL
MICROBIOLOGY,
UCH
JULY 23rd, 2009
 INTRODUCTION
• Genetics is the science that defines and
analyzes heredity, or constancy and
change in the vast array of physiologic
functions that form the properties of
organisms.
• Gene is the unit of heredity, a segment of
DNA that carries in its nucleotide sequence
information for a specific biochemical or
physiologic property.
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 INTRODUCTION
• Genetics is the basis of expression of certain
traits e.g. certain metabolic pathways in
microbes.
• If metabolism is the basis of life, and genes
encode the enzymes THEN genetics is the
basis of microbial life.
• Like in metabolism, many drugs used to treat
infections interfere with microbial DNA
replication or protein synthesis.
• DNA forms the basis of many new diagnostic
tests for bacterial and viral infections
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 INTRODUCTION
• Microbial Genetics is based largely upon
observation of growth.
• Phenotypic variation has been observed
on the basis of a gene's capacity to permit
growth under conditions of selection;
• eg, a bacterial capacity to grow in the
presence of an antibiotic – bacterial drug
resistance.

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DEFINITIONS OF SOME TERMS
• Genome: A cells complete genetic information (usually DNA)

• Chromosomes: structures found in the nucleus of cells,

arranged in pairs, are carriers of hereditary factors/characters
called genes

• Genes: segments of DNA (generally) that code for

functional products

• Plasmids are small genetic elements capable of independent

replication in bacteria and yeasts.

• Episome - a plasmid that can integrate into the chromosome

• Genotype: the genes of anYuguda

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S (potential properties) 5
DEFINITIONS OF SOME TERMS
• Phenotype: actual expressed properties ie the
manifestation of genotype
• Replicons are DNA circles (chromosome and
plasmid), which contain genetic information
necessary for their own replication.
• Transposons are genetic elements that contain
several kbp of DNA, including the information
necessary for their migration from one genetic
locus to another.
• In doing so, they create insertion mutations via
insertion elements.
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 ORGANIZATION OF GENES
• Genetic information is stored as a sequence of
bases in deoxyribonucleic acid (DNA).
• In RNA bacteriophages [eg, Q, MS2] and some
RNA viruses [eg, influenza and reovirus], genetic
information is stored as a sequence of bases in
ribonucleic acid [RNA].
• DNA mostly exist as double strands with
complimentary bases (A-T, G-C).
• The orientation of the double strands is described
as antiparallel – one strand runs in 5’ to 3’
direction and the other in 3’ to 5’.
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 DNA
• The length of a DNA molecule is usually
expressed in thousands of base pairs, or kilobase
pairs (kbp).
• A small virus may contain a single DNA molecule
of 5 kbp, whereas the single DNA molecule that
forms the Escherichia coli chromosome is 4639
kbp.
• Each base pair is separated from the next by
about 0.34 nm, so that the total length of the E
coli chromosome is roughly 1 mm.
• Bacterial DNA structure is described as
Supercoiled and circular.

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 RNA
• Ribonucleic acid (RNA) most frequently
occurs in single-stranded form.
• The base uracil (U) serves in RNA the
hybridization function that thymine (T)
serves in DNA, so the complementary
bases of RNA are A-U and C-G.

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 RNA
• The main function of RNA is communication of
DNA gene sequences in the form of messenger
RNA (mRNA) to ribosomes.
• The ribosomes, which contain ribosomal RNA
(rRNA) and proteins, translate this message into
the primary structure of proteins via aminoacyl-
transfer RNAs (tRNAs).
• Bacterial ribosomes contain three kinds of rRNA
with respective sizes of 120(5S), 1540(16S), and
2900(23S) bases and a number of proteins – 50S
and 30S proteins.

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 BACTERIAL GENOME
• More than 340 microbial genomes have been
sequenced.
• ~ 90% of these genomes consist of a single
circular DNA molecule containing from 580
kbp to more than 5220 kbp of DNA.
• A few bacteria (eg, Brucella species) have
two circular DNA molecules.
• Lack nuclear membrane and are usually
haploid.
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 BACTERIAL GENOME
• Chromosomes and plasmids carries bacterial
genes.
• Chromosomes carries genes essential for bacterial
growth.
• Plasmids carries genes for specialized functions
including those that mediate their transfer from
one organism to another as well as other genes
associated with acquisition or rearrangement of
DNA.
• R Factors - confer drug resistance
• Bacteriocins - proteins toxic to other bacteria
and human cell

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 Examples of Metabolic Activities
Determined by Plasmids
Organism Activity

Pseudomonas species  Degradation of camphor, toluene, octane,

salicylic acid
Escherichia coli  Sucrose uptake and metabolism,
citrate uptake
Klebsiella species  Nitrogen fixation

Streptococcus (group N) Lactose utilization,

galactose phosphotransferase system,
citrate metabolism
Bacillus α-Amylase
stearothermophilus 
Flavobacterium species  Nylon degradation
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• Transposons; these are DNA segments that
can be
inserted into a DNA molecules.
• ie they carry genetic information required for
their
transfer from one chromosome to another.
• They lack genes necessary for their replication.
• They carry genes for transfer of antibiotic
resistance.

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 DNA Replication
• DNA is copied by DNA polymerase
• - Begins at the replication fork
• – In the 5′ → 3′ direction
• – Initiated by an RNA primer
• – Leading strand synthesized continuously
• – Lagging strand synthesized discontinuously
• – Okazaki fragments
• – RNA primers are removed and Okazaki
fragments joined by a DNA polymerase and
DNA ligase
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 DNA Replication

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 Some Enzymes Involved in DNA
Replication and Their Functions
Helicase • Unzip DNA helix
Primase • Synthesises RNA primer
DNA polymerase III • Add bases to the new
DNA chain
• Proofreads the chains for
mistakes
DNA polymerase I • Removes primers
• Closes gaps
Ligase • Final binding of nicks in
DNA during synthesis and
repair
Gyrase • Supercoiling of the DNA

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 DNA Transcription to RNA
• DNA is transcribed to make RNA (mRNA, tRNA,
and rRNA)
• RNA nucleotides: base pairing using DNA as
template
• Transcription begins when RNA polymerase
(enzyme) binds to the promotor sequence
• Transcription proceeds in the 5′ → 3′ direction
• Transcription stops when it reaches the
terminator sequence

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 What are the possible products
from transcription?
• Messenger RNA (mRNA)
• Transfer RNA (tRNA)
• Ribosomal RNA (rRNA)

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 Translation: RNA to protein
• Requrements;
– mRNA
– Ribosomes
– Amino acids
– tRNA

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 Translation
• mRNA is translated in codons
(3 nucleotides)
• Translation of mRNA begins at the start
codon: AUG which codes for metheonine
• Translation ends at a STOP codon: UAA,
UAG, UGA.

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 The Genetic code

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 Translation

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• Bacterial cells can translate mRNA
molecules into proteins as they are being
transcribed
• Allows rapid response of protein
production in response to changing
conditions

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 Genetic Exchange in Bacteria
• Transformation
• Transduction
• Conjugation

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 Transformation
• Gene transfer resulting from the uptake of
DNA from a donor.
• Factors affecting transformation
• DNA size and state
• Sensitive to nucleases
• Competence of the recipient (Bacillus,
Haemophilus, Neisseria, Streptococcus,
pseudomonads spp)
• Competence factors
• Induced competence

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 Transformation
• Steps
– Uptake of DNA
• Gram +
• Gram -
– Recombination
• Legitimate, homologous or general
• recA, recB and recC genes

• Significance
– Phase variation in Neiseseria
– Recombinant DNA technology
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 Transduction
• Bacterial DNA is transferred via a virus
that infects bacteria - bacteriophage
• Resistant to environmental nucleases
• Virulent phages - cause infection and end in
lysis of cell – lytic cycle
• Temperate phage - incorporated into host DNA
• Repressor that prevents destruction of host
DNA - lysogenic cycle
• Commonly seen in staph aureus.

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 Transduction
• Types;
- Generalised – any donor genes transfered
- Specialised – only certain portion of the
donor gene is transfered
• Phases;
- Infection of Donor
- Phage replication and degradation of host DNA
- Assembly of phages particles
- Release of phage
- Infection of recipient
- Homologous recombination
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 Conjugation
• First described in 1946, Edward Tatum and
Joshua Lederberg
• Mediated by one kind of plasmid - F plasmid
• Differs from transformation
– Cells must be of opposite mating types
– Requires direct cell contact
– Transfers larger quantities of DNA
• Donor cells must have plasmid

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 Tatum and Lederberg Expirament: sexual recombination in
bacteria.
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 Conjugation
• Hfr and F-cell
• Replication of Hfr begins in middle of
integrated F factor
• Small piece leads the chromosome into
F-cell
• Donor DNA can recombine (DNA not
integrated is degraded)
• Used to map genes on chromosome
• After 8 minutes, 1 gene transferred
• After 120 min all transferred
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 Conjugation
• Significance
– Gram -ve bacteria
• Antibiotic resistance – R factors
• Rapid spread
– Gram +ve bacteria
• Production of adhesive material by donor
cells

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 Exchange of Genetic Information

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 MUTATIONS
• Causes of mutations in bacteria;
• Most are spontaneous
• Errors made by DNA Polymerase
• UV light exposure
• Mutations may be neutral, beneficial, or
harmful
• Mutagen: Agent that causes mutations
• Spontaneous mutations: Occur in the
absence of a mutagen

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 Types of Mutations
• Base-pair mutation
– Missense mutation
– Nonsense mutation
• Frameshift
– Changes the reading frame

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 MUTATIONS
• Base substitution
(point mutation)→ Change in one
base
• Missense mutation → Result in change in
amino acid
• Nonsense mutation → Results in a
nonsense codon

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 MUTATIONS
• Frameshift mutation → Insertion or
deletion of one or
more nucleotide
pairs.

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 APPLICATIONS
• Genetic Engineering Was Born from Genetic
Recombination
• Genetic engineering involves changing the
genetic material in an organism to alter its
traits or products
• A recombinant DNA molecule contains DNA
fragments spliced together from 2 or more
organisms
• This was made possible with the Discovery of
endonucleases Plasmids and sticky ends
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 Modern applications of Genetic
Engineering
• Pharmaceutical production
–Insulin, interferon, hormones,
vaccines etc.
• Genetically engineered plants
• Animal gene alterations
• Gene probes
• DNA fingerprinting
• The human genome initiative

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 CONCLUSION
• The study of microbial genetics is very
important in that it forms the bases of
genetic engineering more especially the
development of news drugs as well as the
study of bacterial drug resistance.

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 THANK YOU

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