Lecture # 38

Dr. Buckhaults
Exchange of Genetic Information
 Mutations in Bacteria
• Mutations arise in bacterial populations
– Induced
– Spontaneous
• Rare mutations are expressed
– Bacteria are haploid
– Rapid growth rate
• Selective advantage enriches for mutants
• Gene transfer occurs in bacteria
 General Features of
Gene Transfer in Bacteria

• Unidirectional
– Donor to recipient
• Donor does not give an entire chromosome
– Merozygotes
• Gene transfer can occur between species
 Transformation

• Definition: 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)
• Competence factors
• Induced competence
 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
 Transduction

• Definition: Gene transfer from a donor to a

recipient by way of a bacteriophage

• Bacteriophage (phage): A virus that infects

bcteria
 Phage Composition and Structure
• Composition
– Nucleic acid
Head/Capsid
• Genome size
• Modified bases
– Protein
• Protection
• Infection Contractile Tail
Sheath
• Structure (T4) Tail Fibers
– Size (80 X 100 nm)
Base Plate
– Head or capsid
– Tail
 Infection of Host Cells by Phages
• Adsorption
–Tail fibers
– Receptor is LPS for T4
• Irreversible attachment
– Base plate
• Sheath Contraction
• Nucleic acid injection
• DNA uptake
Microbe Library, American Society for Microbiology
www.microbelibrary.org
 Types of Bacteriophage

• Lytic or virulent – Phage that multiply within the

host cell, lyse the cell and release progeny phage
(e.g. T4)
• Lysogenic or temperate phage: Phage that can
either multiply via the lytic cycle or enter a
quiescent state in the bacterial cell. (e.g., )
– Expression of most phage genes repressed
– Prophage – Phage DNA in the quiescent state
– Lysogen – Bacteria harboring a prophage
 Events Leading to Lysogeny
• Circularization of the phage chromosome
– Cohesive ends

Cohesive Ends

Ligase

Linear Double Stranded Opened Circle Closed Circle

 Events Leading to Lysogeny

• Site-specific
recombination
– Phage coded
enzyme gal bio

• Repression of the
phage genome
gal
– Repressor protein
bio
– Specific
– Immunity to
superinfection gal bio
 Termination of Lysogeny
• Induction
gal bio
– Adverse conditions
• Role of proteases gal bio

– recA protein
– Destruction of gal

repressor bio

• Gene expression
• Excision
• Lytic growth bio
gal
 Transduction

• Definition: Gene transfer from a donor to a

recipient by way of a bacteriophage
• Resistant to environmental nucleases
 Transduction
• Types of transduction
– Generalized - Transduction in which potentially
any donor bacterial gene can be transferred
 Generalized Transduction
• Infection of Donor
• Phage replication and degradation of host DNA
• Assembly of phages particles
• Release of phage
• Infection of recipient
• Homologous recombination

Potentially any donor gene can be transferred

 Transduction
• Types of transduction
– Generalized - Transduction in which potentially
any dornor bacterial gene can be transferred.
– Specialized - Transduction in which only
certain donor genes can be transferred
 Specialized Transduction
Lysogenic Phage
• Excision of the gal bio
prophage
• Replication and
release of phage
• Infection of the gal gal
recipient bio bio
• Lysogenization
of the recipient
– Homologous
recombination
also possible
gal
gal bio bio
 Transduction

• Definition
• Types of transduction
• Significance
– Common in Gram+ bacteria
– Lysogenic (phage) conversion
• e.g. Corynebacterium diptheriae toxin
– Toxin derived from lysogenic phage
 Conjugation

• Definition: Gene transfer from a

donor to a recipient by direct
physical contact between cells
• Mating types in bacteria
Donor
– Donor
• F factor (Fertility factor)
– F (sex) pilus
– Recipient
• Lacks an F factor
Recipient
 Physiological States of F Factor

• Autonomous (F+)
– Characteristics of F+ x F- crosses
• F- becomes F+ while F+ remains F+
• Low transfer of donor chromosomal
genes

F+
 Physiological States of F Factor
Integrated (Hfr)
(High Frequency of Recombination)

– Characteristics of
Hfr x F- crosses
• F- rarely becomes
Hfr while Hfr
remains Hfr
• High transfer of F+ Hfr
certain donor
chromosomal genes
 Physiological States of F Factor
• Autonomous with
donor genes (F’)
– Characteristics of F’
x F- crosses
• F- becomes F’
while F’ remains
F’
• High transfer of
donor genes on F’
and low transfer Hfr F’
of other donor
chromosomal
genes
 Mechanism of F+ x F- Crosses

• Pair formation
– Conjugation
bridge
• DNA transfer F+ F- F+ F-
– Origin of
transfer
– Rolling circle
replication
F+ F+ F+ F+
 Mechanism of Hfr x F- Crosses

• Pair formation
– Conjugation
bridge
Hfr F- Hfr F-
• DNA transfer
– Origin of transfer
– Rolling circle
replication
• Homologous Hfr F- Hfr F-
recombination
Microbe Library, American Society for Microbiology
www.microbelibrary.org
 Mechanism of F’ x F- Crosses

• Pair formation
– Conjugation
bridge
F’ F- F’ F-
• DNA transfer
– Origin of transfer
– Rolling circle
replication
F’ F’ F’ F’
 Conjugation
• Significance
– Gram - bacteria
• Antibiotic resistance
• Exponential increase under selective pressure
– Gram + bacteria
• Production of adhesive material by donor cells
 Transposable Genetic Elements

• Definition: Segments of DNA that are able

to move from one location to another
• Properties
– “Random” movement
– Not capable of self replication (not a replicon)
– Transposition mediated by site-specific recombination
• Transposase
– Transposition may be accompanied by duplication
 Types of Transposable Genetic
Elements
• Insertion sequences (IS)
– Definition: Elements that carry no other genes
except those involved in transposition
– Nomenclature - IS1
– Structure (flanking inverted repeats)
– Importance ABCDEFG Transposase GFEDCBA

• Insertional Mutation
•Plasmid insertion
•Phase variation
Phase Variation in Salmonella H Antigens

H1 gene IS H2 gene

H1 H2
flagella flagella
 Types of Transposable Genetic
Elements
• Transposons (Tn)
– Definition: Elements that carry other genes in
addition to those involved in transposition
– Nomenclature - Tn10
– Structure
IS Resistance Gene(s) IS
• Composite Tns
– Importance IS Resistance Gene(s) IS

• Antibiotic resistance
 Plasmids

• Definition: Extrachromosomal genetic

elements that are capable of autonomous
replication (replicon)
• Episome - a plasmid that can integrate into
the chromosome
 Classification of Plasmids

• Transfer properties
– Conjugative
– Nonconjugative
• Phenotypic effects
– Fertility
– Bacteriocinogenic plasmid
– Resistance plasmid (R factors)
 Structure of R Factors

• RTF RTF
– Conjugative
plasmid
Tn 9
– Transfer genes 0

Tn 21
n 1
Tn 8
T
• R determinant
– Resistance genes
– Transposons R determinant