GENETICS OF BACTERIA

&
THEIR VIRUSES
CONTENT
Bacterial & viral genomes
• E. coli
• Bacteriophages & retrovirus

Mechanisms of genetic exchange in bacteria

• Transformation
• Conjugation
• Transduction
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BACTERIA AND VIRUSES
 Small size
 Rapid reproduction
 Selective media (e.g., antibiotics)
 Simple structures and physiology
 Genetic variability
 Complete genome sequences

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 BACTERIA AND VIRUSES
 Their small size, short generation time, and simple
structures have made bacteria and viruses valuable
model systems for genetic studies.
 Many basic concepts of genetics were first deduced from
studies of bacteria and viruses.

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 BACTERIAL GENOME
 One main circular chromosome
with a few thousand genes.
 Variable number of plasmids and
episomes.
 Episomes are similar to plasmids, but
episomes can replicate either
autonomously or as a part of the main
chromosome – in an integrated state
 Asexual reproduction by simple
fission.
 Parasexual processes occur. 6
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PHENOTYPES IN BACTERIA
 Colony color and morphology
 Mutation can change both colony color &
morphology
 Some mutations alter the morphology of
the bacterium without changing colony
morphology
 Nutritional mutants for energy sources
 Wild-type E. coli: phenotype Lac+
genotype lac+
 Mutant type: phenotype Lac-
genotype lac-

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PHENOTYPES IN BACTERIA
 Prototrophs and auxotrophs
 Wild type bacteria called prototrophs can grow on minimal
medium (energy source & some inorganic salts) and can
synthesize all of the metabolites (amino acids, vitamins, purines,
pyrimidines)
 Mutants called auxotrophs will grow if the metabolites is added
to the medium
 Drug and antibiotic resistance
 Wild –type E. coli cells are killed by antibiotics
 Mutants can resist to antibiotics

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ESCHERICHIA COLI
 Escherichia coli (also known as E. coli) is a gram-negative,
facultative anaerobic, rod-shaped and found in the lower
intestine of warm-blooded organisms (endotherms)
 Simple cell structure included only one chromosomal DNA
and a plasmid
 Perform complicated metabolism to maintain its cell growth
and cell division

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 PLASMIDS
 A small DNA molecule within a cell
that is physically separated from
a chromosomal DNA and can
replicate independently.
 Most commonly found as small
circular, double-stranded DNA
molecules in bacteria.
 Often carry genes that may benefit
the survival of the organism, for
example antibiotic resistance.

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PLASMID STRUCTURE

 Consist of
 An origin of replication
 An antibiotic resistance gene
 Restriction enzyme
recognition sites
 The size of plasmids varies
from 1 to over 400 kbp

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THE GENETICS OF VIRUSES
 A virus is a noncellular infectious particle consisting of
nucleic acid (DNA/RNA with single-stranded or double-
stranded) and protein
 A virus is smaller than any cell and has no metabolic
machinery of its own – it has no ribosomes or other
metabolic machinery and it cannot make ATP
 A virus can replicate only in a living cell – to replicate, a virus
must insert its genetic material into a cell of a specific host
 Some viruses are disease-causing agents (pathogens)
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THE GENETICS OF VIRUSES
 Bacteriophages are viruses that infect bacteria
 Virulent phages: reproduce through the lytic cycle, and always
kill the host cells.
 Temperate phages: inactive prophage – phage DNA integrates
into bacterial chromosomes.

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Table 20-1 p326

BACTERIOPHAGE T4
 Double-stranded DNA genome
 Protein head
 Genome contains 168,800 base
pairs and 150 characterized
genes
 Lytic phage

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 STRUCTURE OF BACTERIOPHAGE T4
 Head: hold DNA genome (capsid)
 Tail: pass nucleic acid into host
cell
 Tail fiber: help tail attach to host
cell and recognizing host cell
surface receptors
 The structure of the 6 megadalton
T4 baseplate that comprises 127
polypeptide chains of 13 different
proteins

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WHAT HAPPEN IN THE HOST CELL?

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 BACTERIOPHAGE LAMBDA ()

 Double-stranded DNA genome

 Genome contains 48,502 base
pairs and about 50 genes
 May be lytic or lysogenic

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WHAT HAPPEN IN THE HOST CELL?

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The steps of the lytic cycle are:
 Attachment: Proteins in the "tail" of the
phage bind to a specific receptor on the
surface of the bacterial cell.
 Entry: The phage injects its double-
stranded DNA genome into the
cytoplasm of the bacterium.
 DNA copying and protein synthesis:
Phage DNA is copied, and phage genes
are expressed to make proteins, such as
capsid proteins.
 Assembly of new phage: Capsids
assemble from the capsid proteins and
are stuffed with DNA to make lots of new
phage particles.
 Lysis: The phages then produce an
enzyme that breaks open the host cell,
releasing the new phages
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The lysogenic cycle begins like the lytic
cycle:
 The phage binds to the bacterium at the
attachment sites
 The phage enters the host cell
 The phage DNA integrates into the
bacterial chromosomes by covalent bond
and it remains as inactive prophage
 The prophage is replicated along with
the bacterial DNA and passed on when
the bacterium divides
 Certain stimuli can cause the prophage
to dissociate from the bacterial
chromosome and enter into lytic cycle,
producing new phage particle and lysing
the cell.
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 LYTIC CYCLE VS LYSOGENIC CYCLE
 Lytic cycle is a type of a viral reproduction  Lysogenic cycle is a viral reproduction
mechanism, which results in the lysis of the mechanism where the viral DNA is
infected cell integrated into the host genome
 Viral DNA does not integrate into the host DNA  Viral DNA integrates into the host DNA

 Does not have the prophage state  Have the prophage state

 Host DNA is hydrolyzed  Host DNA is not hydrolysed

 Viral DNA replication occur independently  Viral DNA replication occurs along with the
from the host’s DNA replication host’s DNA replication
 Productivity of viral DNA is high  Productivity of viral DNA is lower

 Short period of time  Take time longer

 Does not allow genetic recombination in the  Allow genetic recombination in the host
host bacterium bacterium

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 RNA VIRUS
 Retrovirus: RNA viruses
that have been integrated
into the host genome
 Reverse transcriptase:
synthesizing DNA from
RNA or DNA template
 HIV and AIDS

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CONTENT
Bacterial & viral genomes
• E. coli
• Bacteriophages & retrovirus

Mechanisms of genetic exchange in bacteria

• Transformation
• Conjugation
• Transduction
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 TERMS OF TRANSFORMATION

 Transformation
The process that cell directly take up the naked genetic material
from its surroundings environment through the cell membrane.
 Competence
The ability of cell taking up DNA across its plasma membrane from
the environment.
 Transformants
The cells that receive genetic material through transformation
 Cotransformed: cells that are transformed by two or more genes
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PROCESS OF TRANSFORMATION
 Key event: A bacterium takes up DNA from the medium in
which it is growing.
 Recombination may take place between the introduced genes
and those of the bacterial chromosome.

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 INDUCTION OF TRANSFORMATION

 Cells that take up DNA through their membranes are said to be

competent.
 Competency is influenced by growth stage, DNA concentration
in the environment, etc.
 The DNA taken up by competent cells virtually can be any type
of DNA under the appropriate conditions.
 Have developed strains of bacteria that are more competent
than wild-type cells.

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 INDUCTION OF TRANSFORMATION

 Inducing factors:
 Treatment with calcium chloride,
 Heat shock,
 An electrical field.
 Bacterial membrane more porous and permeable to DNA
 The efficiency of transformation can also be increased by using
high concentrations of DNA

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 CONSEQUENCE OF TRANSFORMATION

 Foreign DNA integrates into

bacterial genome or replaces
original DNA segment, and then
expresses characteristics
caused by exogenous DNA.
 Contribute to the genetics
variant and evolution of
virulence factors.

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 TERMS OF CONJUGATION
 Conjugation
DNA is transferred between bacteria through a pillus, one-way
traffic from donor cells to recipient cells.
 F factor
A segment of DNA, which contains genes that allow the DNA to
be transferred between cells.
 Sex pilus – connection tube
 Merozygotes
Partial diploid bacterial cells containing F plasmid carrying
some bacterial genes
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 TERMS OF CONJUGATION
 F+ cells
Donor cells containing F factor
 F− cells
Recipient cells lacking F factor
 Hfr cells (high-frequency strains)
Donor cells with F factor integrated into the donor bacterial
chromosome
 F prime (F′) cells
Cells contain F plasmid carrying some bacterial genes
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THE F FACTOR IN E. COLI

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 PROCESS OF CONJUGATION BETWEEN F+ & F-
1. Coming together of donor and recipient cells.
2. F plasmid is transferred to the recipient cell.
3. Rolling-circle replication in the donor cell and ds-DNA
synthesis in the recipient cell.

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 PROCESS OF CONJUGATION BETWEEN HFR & F-
1. F factor is integrated in Bacterial chromosome.
2. Coming together of the donor cell and the recipient cell.
3. F plasmid is transferred to the recipient cell
4. Rolling-circle replication in the donor cell and ds-DNA synthesis
in the recipient cell.
5. Homologous recombination
6. Non-recombinant DNA fragment is degraded by nuclease
enzyme

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 PROCESS OF CONJUGATION BETWEEN F’ & F-
1. Crossing over within Hfr cell
2. Excision of F factor F’ cell
3. Conjugation with F- cell
4. Only part of recipient cell is diploid partial diploid

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 CONSEQUENCE OF CONJUGATION

Natural gene transfer and antibiotic resistance

 Antibiotic resistance comes from the actions of genes located on
R plasmids that can be transferred naturally.
 R plasmids have evolved in the past 60 years since the beginning
of widespread use of antibiotics.
 The transfer of R plasmids is not restricted to bacteria of the
same or even related species.

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 MAPPING BACTERIAL GENES WITH
INTERRUPTED CONJUGATION
 Principle: chromosome transfer always begins within the
integrated F factor and proceeds in a continuous direction
 So genes are transferred according to their sequence on the
chromosome
 The time required for individual genes to be transferred indicates
their relative positions on the chromosome

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 MAPPING BACTERIAL GENES WITH
INTERRUPTED CONJUGATION
 In most genetic maps, unit: distances-percent recombination
 In bacterial maps constructed with interrupted conjugation, the
basic unit of distance is a minute.
 The frequency of gene transfer from donor to recipient cells
decreased for the more distant genes.
 90% of the recipients received the azir allele, but only about 30%
received the gal+ allele

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 TERMS OF TRANSDUCTION
 Transduction
A bacteriophage transfers DNA from a donor cell to a recipient cell.
 Generalized transduction
A random fragment of bacterial DNA is packaged in the phage head
in place of the phage DNA.
 Specialized transduction
Recombination between the phage chromosome and the host
chromosome produces a phage chromosome containing a piece of
bacterial DNA.
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TERMS OF TRANSDUCTION
 Transducing phage
Phage contains a piece of
bacterial DNA inside the
phage coat.
Transducing phage (virus)
 Transductant
Bacterial cell that
received genes from Transduc
another bacterium tant
through transduction

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PROCESS OF GENERALIZED TRANSDUCTION

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PROCESS OF SPECIALIZED TRANSDUCTION

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SIGNIFICANCE OF GENETIC EXCHANGE IN BACTERIA
 Genetic exchange is as important in bacteria as it is in
other organisms.
 Mutation is the source of new genetic variation.
 Recombination produces new combinations of allele.
 Transformation, conjugation, and transduction generate
new combinations of genes in bacteria to allow bacteria to
adapt to new environments.
 Parasexual recombination mechanisms produce new
combinations of genes in bacteria.
 Parasexual mechanisms enhance the ability of bacteria to
adapt to changes in the environment.
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 Transformation Conjugation Transduction
DNA transmitting Absorb foreign One living cells Via bacteriophage
approach DNA (i. e. from transfer DNA
dead cells) directly to the
another
Occurrence In any species (with Mostly within In specific host and
lab techniques’ population close related
help) species
Length of Variable Variable Depend on type of
transferred phage
genetic segments
Factors that affect size of DNA, Can only occur Host species,
the occurrence of nuclease in the If bacteria have F+ nuclease,
the process environment, plasmid integrated site, size
competency of the of DNA
recipient, ect 77
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BACKGROUND READING
 Pierce, B.A., 2012. Genetics: A Conceptual Approach, 4th Ed.,
Chapter 8, pp. 203-237
 Snustad & Simmons, 2009. Principles of Genetics, 5th Ed.,
Chapter 8, pp. 177-203

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