Microbial genetics

Edet E. Udo PhD

Department of Microbiology
Faculty of Medicine. Kuwait University.
Kuwait.
 Microbial genetics
• Objectives:
• Define genetics and heredity
• Discuss the mechanisms of genetic exchange
in bacteria
• Discuss mutation and mutagenic agents
• Discuss the concepts and application of
genetic engineering in Medicine
Definitions
• 1. Genetics:
– is the science of the study of heredity. It includes
the the study of gene replication and transmission.
• 2. Gene :
– a segment of DNA that codes for a functional
product or a linear sequence of DNA that forms a
functional unit of a chromosome.
– The location of a characteristic is the Locus.
– Genes with different information at the
same locus are alleles
• 3. Genotype:
– the genetic composition of an organism- its entire
DNA
• 4. Phenotype:
– the expression of the genes– the proteins and the
properties they confer on the organism
 Bacterial DNA

• Bacterial DNA:

• DNA stores information

used to guide the
replications of DNA in
preparation for cell
division

• 1. Chromosomal
DNA,
• 2. Extra chromosomal
DNA----Plasmid
DNA
 Bacterial DNA

• Bacterial DNA Replication:

– by the semi conservative mode,
– Replication of DNA usually begins at a specific
point origin of replication)
– During binary fission each daughter cell
receives a chromosomal DNA like the one in the
parent cell.
 Mutations
• Definition:
• Mutations are alterations
(changes) in the nucleic
acid sequence.
– accounts for variations
in the genotypes and
phenotypes of
microorganisms
Types of mutations :
• Point mutations:
consists of changes in a
single nucleotide
• Frameshift
mutations: consists of
of the insertion or deletion
of a single nucleotide
 Mutations
• Effects of mutation:
Phenotypic variations
e.g lethal mutation leading to arrest of protein
synthesis
– No effect:
• Spontaneous mutation:
• Occurs in the absence of known mutagen
• May be due to errors in base pairing
• Induced mutation:
• Produced by agents known as mutagens
• Antibiotic resistance,
• Generation of new phenotypes.
 Mutagens
• Chemical agents:
• 5-bromouracil, nitrous acid, ethidium
bromide, nitroguanidine, acridines, etc.
• Physical agents:
• X-rays, UV-rays: causes formation of dimers
• Biological mutants:
• Transposons, insertion sequences.
 Mutants
• Repair of DNA damage:
• Light repair:
• involves enzymes that is activated by visible
light and breaks bonds between pyrimidines of
a dimer
– ( role in skin cancer)
• Dark repair:
• involves several enzymes that do nor require
light for DNA repair.
• The AMES test:
• Itis used to identify possible chemical
carcinogens in chemicals applied to human body
such as body creams, shampoos etc.
• It is based on the ability of bacteria to mutate by
reverting to their original synthetic ability
(reverse mutation).
• E.g. Histidine auxotroph of Salmonella are
exposed to potential carcinogen and revertants
to the non mutant state shows that the chemical
is a potential carcinogen.
 Genetic transfer and
recombination
• Definition:
– Gene transfer refers to the movement of
genetic information between organisms.
– Occurs by transformation, transduction and
conjugation.
• Transformation:
– involves the uptake of naked DNA by
bacteria. Uptake of DNA .
– It was first demonstrated in 1928 by Griffith
while studying pneumococcal infections in
mice.
– Occurs naturally in Streptococcus
pneumoniae, Heamophilus species and
some Bacillus species
 Transformation

• Transformation
is significant
because:
– It contributes to
genetic diversity
– It can be used to
introduce DNA into
organisms, observe
its effect and study
gene location
– It can be used to
create recombinant
DNA
 Genetic transfer and
recombination
• Transduction:
• mode of transfer involving bacteriophages.
• Significance of transduction.
– Transfers genetic materials and
demonstrates a close evolutionary
relationship between host cell DNA and
prophage.
– Its persistence in a cell suggests a
mechanism for viral origins of cancer.
– Provides a mechanism for studying gene
linkage
• Phages can be virulent (lytic) or temperate.
• 1. Virulent phages
• destroy a host cell’s DNA, and cause lysis of the
host cell in a lytic cycle
• 2. Temperate phages (prophages)
– produces a repressor substance that prevents destruction of
host DNA.
• Cells containing prophages are called Lysogenic
cells because they have the potential to enter the
lytic cycle
Phage cycle
 Transduction
• Transduction can
be specialized or
• generalized.
• Generalized
transduction,
• the phage can
incorporate any part of
the chromosomal or
plasmid DNA and
transfer them.
 Transduction
• In specialized
transduction the
phage is
incorporated into
the chromosome
and can transfer
only genes
adjacent to the
phage.e.g. phage
lambda in E. coli
 Lysogenic or phage conversion
• The alteration of a bacterial
phenotype resulting from the
acquisition of a phage.
• It may confer virulence
property . Examples include:
The production of diphtheria
toxin after the acquisition of
phage B
The acquisition of Shiga-like
toxin by E.coli after acquisition
of a phage
Production of botulinum toxin-
C by phage
Production of scarlet fever
toxin by lysogenic
Streptococcus pyogenes
 Genetic transfer and
recombination
• Conjugation: A process requiring cell to cell contact.
– 1. Contact between donor and recipient cells is
required
– 2. Larger amount of DNA is transferred.
– Mediated by conjugative plasmids or conjugative
transposons.
– Sex pilli is involved In E. coli and other Gram-
negative bacilli,
– Sex pheromones may be involved in Streptococci

• Significance of conjugation:
– It increases genetic diversity
– May represent an evolutionary stage between
asexual and sexual reproduction
– It provides a means of mapping genes in bacterial
chromosome
 Conjugation

Conjugatio
n in E. coli
 Genetic transfer and
recombination
• Characteristics of plasmids:
• Double stranded extra chromosomal DNA.
• Plasmids are self-replicating
• They are identified by virtue of some recognizable
function that they serve in a bacterium e.g. F-plasmids
(fertility factors) direct the synthesis of proteins that
self-assemble into sex pili
• R-plasmids (resistance factors) carry genes that
provide resistance to various antibiotics
• Other plasmids direct the synthesis of bactericidal
proteins called bacteriocin, toxin plasmids ( metabolic
plasmids (Tol plasmids)
 Plasmids
• Classification:
• based on size, copy
number, phenotype,
incompatibility
• Restriction
endonucleases
analysis ( physical
mapping)
• Plasmids are used in
the study of Agarose gel electrophoresis of plasmids.
epidemiology of
pathogenic bacteria.
 Genetic engineering
Definition:
the purposeful manipulation of genetic
material to alter the characteristics of an
organism.
• Techniques include genetic fusion, protoplast
fusion and recombinant DNA.
• Recombinant DNA makes it possible to fuse
genes with vectors and clone them in host
cells.
• Making Recombinant DNA
involves:
• The manipulation of DNA in vitro,
• The cloning of DNA from other
organism s in bacteria DNA with
phage or plasmid.
• The production of many genetically
identical progeny of phages or
plasmids.
 Applications of Recombinant DNA
technology in Medicine
• 1. Treatment or management of disease
– Production of growth hormones, insulin, anti sera, blood
coagulation proteins etc
– Gene therapy
• 2. Diagnosis of disease
– DNA probes for detection of genetic defects, identification of
pathogens
– DNA amplification (PCR)
• 3. Prevention and control of infections
– Recombinant vaccines
– Molecular epidemiology- for the determination of source and
extent of spread of an infectious agent
• Safety concerns:
• Environmental impact, health effect of recombinant plants and
vegetables,
• Production of Biological warfare agents, ethical concern with
use on human subjects