Microbiology

Chapter 2 Microbial Genetics

19-Sep-2020
Mr: Qadar Ahmed Isse
 Mutations
Change in genetic material.
1. Point mutations = base pair substitution (silent, missense,
nonsense)
2. Frameshift mutations = Insertion or deletion of one or
more nucleotide pairs

Review
Fig 8.17
 Various Point Mutations

Missense

Nonsense

Silent
Fig 8.17
 Mutations cont.
• May be neutral (silent), beneficial, or harmful.
• Spontaneous mutation rate  10-6  1 mutation per
million replicated genes
• Mutagens increase mutation rate 10 – 1000x
Chemical mutagens
– Nucleoside (base) analogs have altered base-pairing
properties. They can be
• randomly incorporated into growing cells (cancer drugs)
• only used by viral enzymes (e.g. AZT)
– Frameshift mutagens such as intercalating agents (e.g.:,
aflatoxin, ethidium bromide)
Fig 8.19a
Distortion due to
intercalating agent
will lead to one or
more base-pairs
inserted or deleted
during replication.

Potent carcinogens!
 Radiation as a Mutagen

1. Ionizing radiation (x-rays and -rays)

lead to deletion mutations (ds breaks)
2. UV rays lead to thymine
dimers (intrastrand bonding)
– Photolyases = light repair enzymes
(use energy from visible light to fix UV light damage)
– Nucleotide excision repair for repair of
all mutations
 Repair
• Photolyases separate thymine dimers
• Nucleotide excision repair

Fig 8.20

ANIMATION Mutations: Repair

 Mutagen Identification: Ames Test
• Wild type vs. mutant
• Auxotroph vs. prototroph
• Many mutagens are carcinogens

Combine animal liver cell extracts with Salmonella

auxotroph
 Expose mixture to test substance
 Examine for signs of mutation in Salmonella,
i.e. Look for cells (colonies) that have reverted
from his– to his+
• Auxotroph VS Prototroph
• Prototroph: Micro-organism that has same
nutritional requirements as the parent organism
• Auxotroph: an organism or cell capable of
synthesizing its all metabolites from inorganic
material requiring no Organic Nutrients.
Ames Reverse Gene Mutation Test

Fig. 8.22
Professor Richard A. Muller of UC Berkeley
on the Ames Test and Natural Foods

Positive or
negative
Ames
test?

Explain
what
happened
 Genetic Transfer and Recombination

• Vertical gene transfer: Occurs during

reproduction between generations of cells.
• Horizontal (lateral) gene transfer: Transfer of
genes between cells of the same generation.
Leads to genetic recombination
• Three mechanisms of horizontal gene transfer:
– Transformation
– Conjugation
– Transduction
 Genetic Recombination
Vertical gene transfer: Occurs during
reproduction between generations
of cells.
Horizontal gene transfer: The transfer
of genes between cells of the same
generation. Leads to genetic
recombination.
Three mechanisms of horizontal
gene transfer:
1. Transformation
2. Conjugation
3. Transduction
 1) Transformation
“Naked” DNA transfer

Recipient cells have to be “competent”

Occurs naturally among very few genera (G+ and G)

Simple laboratory treatment will make E. coli

competent  workhorse for genetic engineering

Griffith’s historical experiment in 1928

Griffith’s Experiment to Demonstrate
Genetic Transformation
Fig 8.24
Transformation
and
Recombination

Fig 8.25
 2) Conjugation
Plasmid and chromosomal DNA
transfer via direct cell to cell contact
High efficiency
F+ = donor cell. Contains F plasmid
(factor) and produces conjugation
(F) pilus (aka “sex pilus”)
Recipient cell (F– ) becomes F+
In some cells F factor integrates into chromosome
 Hfr cell
R plasmids (R factors) are also transferred via
conjugation
ANIMATIONs
Fig 8.27
 3) Transduction
DNA Transfer from donor to
recipient cell with help of
bacteriophage (= transducing phage)

2 types of phage-bacteria interaction:

1. Generalized transduction happens via
lytic cycle caused by virulent phages

2. Specialized transduction happens via

lysogenic cycle of a temperate
bacteriophage
Transduction by a
Bacteriophage

ANIMATION Generalized
Transduction

ANIMATION Specialized
Transduction

Fig 8.28