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MICROBIAL GENETICS
BY
DR JAWAD NAZIR
ASSISTANT PROFESSOR
DEPARTMENT OF MICROBIOLOGY
UNIVERSITY OF VETERINARY AND ANIMAL SCIENCES, LAHORE
Microbial genetics
Terminology
Genetics Study of what genes are, how they
carry information, how information is
expressed, and how genes are
replicated
Gene Segment of DNA that encodes a
functional product, usually a protein
Microbial genetics
Terminology
Genome
 All of the genetic material in a cell
Genomics
 Molecular study of genomes
Genotype
 Genetic make up of an organism (potential properties)
 Collection of genes
Phenotype
 Expression of the genes
 Collection of proteins
Microbial genetics
Chromosome (E. coli)
• 4.6 million base pairs
• 1 meter long
• 1000 times longer than
bacteria
 Polymer of nucleotides:
adenine, thymine,
cytosine, guanine
 Double helix associated
with proteins
 "Backbone" is
deoxyribose-phosphate
 Strands held together
by hydrogen bonds
between AT and CG
 Strands are antiparallel
DNA
Microbial genetics
 Consist of nucleotides.
 Nucleotides consist of a:
• Pentose
• Phosphate group
• Nitrogen-containing (purine or pyrimidine) base
Nucleic Acids
Microbial genetics
Flow of Genetic Information
DNA replication
Microbial genetics
DNA replication
 Two parent strands unwound
 DNA polymerase
 Replication fork
 Semiconservative replication
 Each parent strand make complementary strand
Microbial genetics
 DNA is copied by DNA polymerase
 In the 5  3 direction
 Initiated by an RNA primer
 Leading strand synthesized continuously
 Lagging strand synthesized discontinuously
 Okazaki fragments
 RNA primers are removed and Okazaki fragments joined
by a DNA polymerase and DNA ligase
DNA replication
DNA replication
Microbial genetics
Adding nucleotides to a DNA
Microbial genetics
DNA replication
Bidirectional
Closed loop DNA
Microbial genetics
 DNA is transcribed to make RNA (mRNA, tRNA,
and rRNA)
 Transcription begins when RNA polymerase binds
to the promoter sequence
 Transcription proceeds in the 5  3 direction
 Transcription stops when it reaches the
terminator sequence
Transcription
Microbial genetics
Transcription
Transcription
Transcription
Transcription
Microbial genetics
 Process of protein synthesis from mRNA strand
 mRNA is translated in codons (3 nucleotides)
 Translation of mRNA begins at the start codon: AUG
 Translation ends at a STOP codon: UAA, UAG, UGA
 Sense codons (61)
 Nonsense codons (3) stop codons
 UAA, UAG, UGA
Translation
Translation
Translation
Translation
Translation
Translation
Translation
Translation
Translation
28
Translation
Translation
Polyribosome
Microbial genetics
RNA processing in prokaryotes
Microbial genetics
RNA processing in eukaryotes
Microbial genetics 33
 Constitutive enzymes are expressed at a fixed rate
 Other enzymes are expressed only as needed
 Repressible enzymes
 Inducible enzymes
 Beta galactosidase in E.coli
 Insulin
Regulation of Bacterial Gene Expression
Microbial genetics
Regulation of Gene Expression
Microbial genetics
Operon
Repression
Regulation of Gene Expression
Regulation of Gene Expression
Regulation of Gene Expression
Regulation of Gene Expression
Microbial genetics 41
 Change in the genetic material
 Mutations may be neutral, beneficial, or harmful
 Mutagen: Agent that causes mutations
 Spontaneous mutations: Occur in the absence of a
mutagen
Mutation
Point mutation
 Base substitution (point
mutation)
 Missense mutation
Mutation
•Change in one base
•Result in change in
amino acid
 Nonsense mutation
Mutation
• Results in a nonsense
codon
 Frameshift mutation
Mutation
•Insertion or deletion of one
or more nucleotide pairs
Microbial genetics 46
Mutagens
 Chemical mutagens
 A number of household and lab chemicals sodium azide
 Physical mutagens
 Radiation, UV light
Mutation
Microbial genetics
Chemical mutagens
Nitrous acid as a mutagen
Nucleoside analogs
Microbial genetics 49
 Ionizing radiation (X rays and gamma rays) causes the
formation of ions that can react with nucleotides and
the deoxyribose-phosphate backbone.
 Breakage of covalent bonds in phosphate sugar
backbone of DNA
 Covalent bonds between adjoining nitrogenous bases
caused by UV light (thymin dimer)
 Nucleotide excision repairs mutations
Radiations
50
 UV radiation causes
thymine dimers
 Light-repair
separates thymine
dimers
 Skin cancer by
accumulation of
TDs in the skin cells
Mutation
Microbial genetics 51
 Spontaneous mutation rate = 1 in 109 replicated
base pairs or 1 in 106 replicated genes
 Mutagens increase to 10–5 or 10–3 per replicated
gene
The Frequency of Mutation
Microbial genetics 52
Identifying mutants
 Bacteria have one copy of each gene so identifying
mutation is relatively easy
Selection:
 Positive (direct) selection detects mutant cells because
they grow or appear different.
 Pencillin resistant population
 Negative (indirect) selection detects mutant cells
because they do not grow.
Replica Plating
The Ames Test for Chemical Carcinogens
Genetic transfer and Recombination
Genetic transfer and Recombination
Microbial genetics 57
 Vertical gene transfer
 Horizontal gene
transfer
Genetic Transfer and Recombination
Occurs during reproduction,
between generations of
cells
Transfer of genes between
cells of the same
generation
58
Transformation
In vitro transformation
Mechanim of transformation
Mechanim of transformation
Microbial genetics
 Competant cells
 Naturally occuring
 Induced
 Calcium chloride
Transformation
Microbial genetics
Conjugation
Microbial genetics
Conjugation
Microbial genetics 65
Conjugation
Microbial genetics 66
Conjugation
Microbial genetics
Conjugation
68
Transduction
Recombinant
1
Phage protein coat
Bacterial
chromosome
2
3
Bacterial
DNA
Phage
DNA
4
Recipient cell
5
Donor
bacterial
DNA
Recipient
bacterial
DNA
Recombinant cell
A phage infects the
donor bacterial
cell.
Phage DNA and proteins
are made, and the
bacterial chromosome is
broken down into pieces.
Occasionally during phage assembly,
pieces of bacterial DNA are packaged
in a phage capsid. Then the donor cell
lyses and releases phage particles
containing bacterial DNA.
A phage carrying bacterial
DNA infects a new host cell,
the recipient cell.
Recombinant can occur,
producing a recombinant
cell with a genotype
different from both the
donor and recipient cells.
Microbial genetics 69
 Conjugative plasmid Carries genes for sex
pili and transfer of the
plasmid
 Dissimilation plasmids Encode enzymes for
catabolism of unusual
compounds
 R factors Encode antibiotic
resistance
Plasmids
Microbial genetics 70
Plasmids
Microbial genetics 71
 Segments of DNA that can move from one region
of DNA to another
 Contain insertion sequences for cutting and
resealing DNA (transposase)
 Complex transposons carry other genes
Transposons
Microbial genetics
Insertion sequence
Microbial genetics
Complex transposons
Insertion of transposons into plasmids
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microbial genetics