Microbiology

Lecture 6: Bacterial Genetics

July 19, 2012
OLFU College of Medicine Class 2015
Dr. Santos
Ut In Omnibus Glorificetur Deus Page 1 of 4




Bacterial Genetics
Plasmid
Autonomous extra-chromosomal elements composed of
circular, double-stranded DNA
Found in most species of gram-positive and gram-negative
bacteria
Carry important genetic info virulence properties like
antibiotic resistance genes
1/20
th
size of a chromosome

Mutation
Any alteration in the determinants of hereditary
characteristics
Heritable change in genome
Changes in DNA sequence

Kinds of Mutation
A. Spontaneous Mutation
Occurs without apparent cause
Occurs with a low frequency of 10-6 to 10-12 in a
population derived from a single bacterium
Ex. Polymerase mistakes

B. Induced Mutation
Caused by mutagens
Increases the frequency of mutation

Effects of Mutation
A. Silent Mutation
Change at the DNA level
Does not result in any change in amino acid in encoded
protein

B. Missense Mutation
Results in substation of one amino acid for another
May be without discernible phenotypic effect
May cause inactivation of the gene product

C. Nonsense Mutation
One which generates a nonsense causing a premature
termination of protein synthesis
Codon encoding an amino acid is changed to a stop codon

D. Frameshift mutation
Small deletion or in insertion that is not in multiples of
three resulting to a change in the reading fram
Lead to premature truncation of protein

E. Null Mutation
Occurs with extensive insertion, deletion or gross
rearrangement of chromosome structure that completely
destroys gene function

Transposable Elements
Genetic units that are capable of mediating their own
transfer: from one chromosome to another, from one
location to another on the same chromosome, between
chromosome and plasmid
This transposition relies on their ability to synthesize
their own specific recombination enzyme Transposase

Characteristics of Transposable Elements
Have inverted complementary repeated sequences of
about 10-40 base pairs at the ends
the DNA surrounding an element always contains a
directly repeated sequences of a small number of base
pairs
these are not part of the element but rather generated
from the target DNA upon insertion of the IS elements







A. Insertion Sequence
Segments of DNA of approximately 600-2000 base pairs
Has single gene that codes for transposase
Has inverted repeats at their termini
Simplest transposable elements bounded by directly
repeated nucleotide sequences (direct repeats, not part of
the IS element)
Can be components of transposons


Figure 1. Insertion Sequence

B. Transposons
Transposable segments of DNA containing genes beyond
those need for transposition
As much as 10-fold larger than IS elements
Has inverted repeats or inverted IS elements at termini
Has transposase gene
Also bounded by direct repeats
Carry other genes (e.g. antimicrobial resistance)

Figure 2. Tranposon

C. Transposable Prophage (Bacteriophage Mu)
During lysogeny, the Mu phage can insert virtually
anywhere in the E. coli chromosome and later can
transpose itself from one location to another
A transposon

Consequences of Transposition
inactivation of a gene into which the element transposes
activation of a gene
transpositional recombination (rearrangement of DNA)
generation of large deletion mutations in bacterial
chromosome
generation of composite transposons from isolated IS
elements and drug resistance genes

Genetic Exchange in Bacteria

A. Transformation
involves the release of DNA into the environment by the
lysis of some cells, followed by the direct uptake of the
DNA by the recipient
requires competence of recipient cells
the transforming factor is usually a double-stranded DNA
occurs among related species
occurs in certain gram-positive and gram-negative
bacteria such as Haemophilus influenza, Streptococcus
pneumonia, Bacillus sp, Neisseria sp.
Used in genetic engineering, recombinant DNA technology




Ut In Omnibus Glorificetur Deus Page 2 of 4

Figure 3. Bacterial Transformation

B. Conjugation
DNA passed directly by cell-to-cell contact during the
mating of the bacteria
Sex-like exchange that requires sex pili
one-way transfer of DNA from a F
+
donor (male) cell with
a sex pilus to a F
-
recipient (female)
unidirectional
mating process is controlled by an F (fertility) plasmid,
which carries the genes for the proteins required for
conjugation
a cell without the F factor is referred to as the F- cell
occurs in E. coli, Bacteroides sp., Streptococci, Streptomyces,
Clostridia
first known conjugation E. coli and Shigella


Figure 4. Bacterial Conjugation

Three States
1. F
+

o Autonomously replicating plasmid
o Transfers only the genes of the F factor
o Mating between the an F+ and an F- cell results in
the acquisition of the maleness by the recipient
(F- to F+)
2. F
o Autonomously replicating plasmid that contains a
small segment of the chromosome
o Transfers a small segment of the chromosome
o Mating between F and F- will result to the
acquisition of the maleness of the recipient



3. Hfr
o High frequency recombinant
o Episome (plasmid integrated with the chromosome)
o Can transfer chromosomal genes
o Mating between Hfr and F- will NOT result in
acquisition of maleness by the recipient
o During this transfer, the single strand of DNA that
enters the recipient F- cell contains a piece of the F
factor
o F plasmid sequence becomes incorporated into the
recipient bacterial chromosome


Figure 5. Hfr Transfer

For helpful videos regarding transformation and conjugation:
http://www.pc.maricopa.edu/Biology/rcotter/Title%205%20Files
/GeneticsLB/GeneticsLB10.html

C.Transduction
Mediated by a bacteriophage (DNA + capsid)
Bacteriophage picks up fragments of DNA and package
them into bacteriophage partciles
DNA is delivered to infected cells and incorporated into
the recipients genome

1. Virulent Phage (Lytic Cycle)
o Causes generalized transduction
o Any part of the host chromosome can be transferred
to the recipient
o Multiplies in the host cell and cause lysis of the host
o Virulent bacteriophages start their life cycle when they
adsorb to a permissive host
o They inject their genetic material into the host and
start to produce viral proteins and copies of the virus
genome, using bacterial resources and biosynthetic
apparatus
o Progeny virus particles are formed and after
completing the cycle, they are released after host cell
lysis
o Only when lysogenized by a particular phage are
bacteria able to produce toxins
o Ex. Lysogenized Group A streptococci scarlet fever
strawberry tongue
o For instructional video, visit
http://www.phageconsultants.com/virulent.ph
p





Ut In Omnibus Glorificetur Deus Page 3 of 4
a. Adsorption during the Lytic Life Cycle of a Lytic
Bacteriophage

The bacteriophage binds to receptors on the bacterial cell wall.
b. Penetration during the Lytic Life Cycle of a Lytic
Bacteriophage

The bacteriophage injects its genome into the bacterium's
cytoplasm.
c. Early Replication during the Lytic Life Cycle of a Lytic
Bacteriophage

The bacteriophage genome replicates and bacteriophage
components begin to be produced by way of the host
bacterium's metabolic machinery.
d. Late Replication during the Lytic Life Cycle of a Lytic
Bacteriophage

The production of bacteriophage components and enzymes
progresses.










e. Maturation during the Lytic Life Cycle of a Lytic
Bacteriophage

The bacteriophage components assemble.
f. Release during the Lytic Life Cycle of a Lytic
Bacteriophage

A bacteriophage-coded enzyme breaks down the
peptidoglycan in the bacterial cell wall causing osmotic lysis.

2. Temperate Phage (Lysogenic Cycle)
o Causes specialized transduction
o Only specific genes are transfereed to the recipient (those
genes that are adjacent to the integration site
o Usually does not multiple upon entry into the host cell but
instead its DNA integrates with the host chromosome,
causing a state of lysogeny

o Temperate phages are basically bacteriophages which can
choose between a lytic and lysogenic pathway of
development.
o Virus remains dormant until induction.
o Temperate bacteriophages start their life cycle when they
adsorb to permissive host.
o After injecting their genome into the host cell, they produce
a set of early proteins and a few copies of their genome.
o On this stage a decision "lysis versus lysogeny" is made.
o Usually in poor growth conditions of the host cell, a phage
chooses lysogenic pathway, because the number of progeny
it can produce in such cell is usually low.
o When lysogeny is chosen, the phage integrates its genetic
material with the host cell.
o It may be done by physical incorporation of the phage
genome into host genome, or the prophage may be
integrated as a stably maintained plasmid.
o When a prophage is induced it starts to produce viral
proteins and copies of virus the genome using bacterial
resources and biosynthetic apparatus.
o Progeny virus particles are formed and, after completing
the cycle, released during host cell lysis.
o For instructional video, visit
http://www.phageconsultants.com/temperate.php


Ut In Omnibus Glorificetur Deus Page 4 of 4