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Lecture 6 - Microbial Genetics

This document discusses microbial genetics and genetic engineering. It defines key terms like genetics, genes, genotype, and phenotype. It describes bacterial DNA structure and replication, as well as mechanisms of genetic exchange in bacteria like transformation, transduction, and conjugation. It discusses mutation, mutagens, and repair of DNA damage. The document also covers plasmids, genetic engineering techniques like recombinant DNA, and applications of recombinant DNA technology in medicine.

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1K views26 pages

Lecture 6 - Microbial Genetics

This document discusses microbial genetics and genetic engineering. It defines key terms like genetics, genes, genotype, and phenotype. It describes bacterial DNA structure and replication, as well as mechanisms of genetic exchange in bacteria like transformation, transduction, and conjugation. It discusses mutation, mutagens, and repair of DNA damage. The document also covers plasmids, genetic engineering techniques like recombinant DNA, and applications of recombinant DNA technology in medicine.

Uploaded by

api-3703352
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
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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

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