Foundations in

Microbiology
Sixth Edition
Chapter 9
Microbial Genetics
Lecture PowerPoint to accompany
Talaro
Copyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display.
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Genetics and Genes
Genetics the study of heredity

The science of genetics explores:
1. Transmission of biological traits from parent
to offspring
2. Expression and variation of those traits
3. Structure and function of genetic material
4. How this material changes
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Levels of Structure and Function of
the Genome
Genome sum total of genetic material of an
organism (chromosomes + mitochondria/chloroplasts
and/or plasmids)
genome of cells DNA
genome of viruses DNA or RNA
DNA complexed with protein constitutes the genetic
material as chromosomes.
Bacterial chromosomes are a single circular loop.
Eucaryotic chromosomes are multiple and linear.
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Chromosome is subdivided into genes, the
fundamental unit of heredity responsible for a
given trait.
site on the chromosome that provides information
for a certain cell function
segment of DNA that contains the necessary code to
make a protein or RNA molecule
Three basic categories of genes:
1. Genes that code for proteins - structural genes
2. Genes that code for RNA
3. Genes that control gene expression - regulatory
genes

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All types of genes constitute the genetic
makeup genotype.
The expression of the genotype creates
observable traits phenotype.
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Genomes Vary in Size
Smallest virus 4-5 genes
E. coli single chromosome containing
4,288 genes; 1 mm; 1,000X longer than cell
Human cell 46 chromosomes containing
31,000 genes; 6 feet; 180,000X longer than
cell

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DNA
Two strands twisted into a helix
Basic unit of DNA structure is a nucleotide
Each nucleotide consists of 3 parts:
a 5 carbon sugar - deoxyribose
a phosphate group
a nitrogenous base adenine, guanine, thymine,
cytosine
Nucleotides covalently bond to form a sugar-
phosphate linkage the backbone
each sugar attaches to two phosphates
5 carbon and 3 carbon
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DNA
Nitrogenous bases covalently bond to the 1
carbon of each sugar and span the center of the
molecule to pair with an appropriate
complementary base on the other strand
adenine binds to thymine with 2 hydrogen bonds
guanine binds to cytosine with 3 hydrogen bonds
Antiparallel strands 3 to 5 and 5 to 3
Each strand provides a template for the exact
copying of a new strand
Order of bases constitutes the DNA code


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Significance of DNA Structure
1. Maintenance of code during reproduction
- constancy of base pairing guarantees
that the code will be retained.
2. Providing variety - order of bases
responsible for unique qualities of each
organism.

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DNA Replication
Making an exact duplicate of the DNA involves
30 different enzymes
Begins at an origin of replication
Helicase unwinds and unzips the DNA double
helix
An RNA primer is synthesized by primase
DNA polymerase III adds nucleotides in a 5 to 3
direction
leading strand synthesized continuously in 5 to 3
direction
lagging strand synthesized 5 to 3 in short segments;
overall direction is 3 to 5


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DNA polymerase I removes the RNA
primers and replaces them with DNA.
When replication forks meet, ligases link
the DNA fragments along the lagging strand
to complete the synthesis.
Separation of the daughter molecules is
complete.
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DNA replication is semiconservative because
each chromosome ends up with one new
strand of DNA and one old strand.
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Applications of the DNA code
Information stored on the DNA molecule is
conveyed to RNA molecules through the
process of transcription.
The information contained in the RNA
molecule is then used to produce proteins in
the process of translation.
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Gene-Protein Connection
1. Each triplet of nucleotides on the RNA
specifies a particular amino acid.
2. A proteins primary structure determines its
shape and function.
3. Proteins determine phenotype. Living things
are what their proteins make them.
4. DNA is mainly a blueprint that tells the cell
which kinds of proteins to make and how to
make them.
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RNAs
Single-stranded molecule made of nucleotides
5 carbon sugar is ribose
4 nitrogen bases adenine, uracil, guanine, cytosine
phosphate
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RNA
3 types of RNA:
messenger RNA (mRNA) carries DNA message
through complementary copy; message is in triplets
called codons
transfer RNA (tRNA) made from DNA;
secondary structure creates loops; bottom loop
exposes a triplet of nucleotides called anticodon
which designates specificity and complements
mRNA; carries specific amino acids to ribosomes
ribosomal RNA (rRNA) component of ribosomes
where protein synthesis occurs



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Transcription
1. RNA polymerase binds to promoter region upstream
of the gene.
2. RNA polymerase adds nucleotides complementary
to the template strand of a segment of DNA in the 5
to 3 direction.
3. Uracil is placed as adenines complement.
4. At termination, RNA polymerase recognizes signals
and releases the transcript.
100-1,200 bases long
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Translation
Ribosomes assemble on the 5 end of a
mRNA transcript.
Ribosome scans the mRNA until it reaches
the start codon, usually AUG.
A tRNA molecule with the complementary
anticodon and methionine amino acid enters
the P site of the ribosome and binds to the
mRNA.

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Translation Elongation
A second tRNA with the complementary
anticodon fills the A site.
A peptide bond is formed.
The first tRNA is released and the ribosome
slides down to the next codon.
Another tRNA fills the A site and a peptide
bond is formed.
This process continues until a stop codon is
encountered.

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Translation Termination
Termination codons UAA, UAG, and
UGA are codons for which there is no
corresponding tRNA.
When this codon is reached, the ribosome
falls off and the last tRNA is removed from
the polypeptide.
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The Master Genetic Code
Represented by the mRNA codons and the
amino acids they specify
Code is universal
Code is redundant


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Polyribosomal complex allows for the synthesis of
many protein molecules simultaneously from the
same mRNA molecule.
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Eucaryotic Transcription and
Translation
1. Do not occur simultaneously transcription
occurs in the nucleus and translation occurs in
the cytoplasm.
2. Eucaryotic start codon is AUG, but it does not
use formyl-methionine.
3. Eucaryotic mRNA encodes a single protein,
unlike bacterial mRNA which encodes many.
4. Eucaryotic DNA contains introns intervening
sequences of noncoding DNA- which have to be
spliced out of the final mRNA transcript.
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Genetics of Animal Viruses
Viral genome - one or more pieces of DNA
or RNA; contains only genes needed for
production of new viruses
Requires access to host cells genetics and
metabolic machinery to instruct the host cell
to synthesize new viral particles

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Regulation of Protein Synthesis and
Metabolism
Genes are regulated to be active only when
their products are required.
In procaryotes this regulation is coordinated
by operons, a set of genes, all of which are
regulated as a single unit.

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Operons
2 types of operons:
inducible operon is turned ON by substrate:
catabolic operons- enzymes needed to metabolize
a nutrient are produced when needed
repressible genes in a series are turned OFF by
the product synthesized; anabolic operon
enzymes used to synthesize an amino acid stop
being produced when they are not needed
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Lactose Operon: Inducible Operon
Made of 3 segments:
1. Regulator- gene that codes for repressor
2. Control locus- composed of promoter and
operator
3. Structural locus- made of 3 genes each coding
for an enzyme needed to catabolize lactose
b-galactosidase hydolyzes lactose
permease - brings lactose across cell membrane
b-galactosidase transacetylase uncertain function
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Lac Operon
Normally off
In the absence of lactose, the repressor binds
with the operator locus and blocks transcription
of downstream structural genes.
Lactose turns the operon on.
Binding of lactose to the repressor protein
changes its shape and causes it to fall off the
operator. RNA polymerase can bind to the
promoter. Structural genes are transcribed.
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Arginine Operon: Repressible
Normally on and will be turned off when
nutrient is no longer needed
When excess arginine is present, it binds to
the repressor and changes it. Then the
repressor binds to the operator and blocks
arginine synthesis.
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Antibiotics That Affect
Transcription and Translation
Rifamycin binds to RNA polymerase
Actinomycin D - binds to DNA and halts
mRNA chain elongation
Erythromycin and spectinomycin interfere
with attachment of mRNA to ribosomes
Chloramphenicol, linomycin and tetracycline-
bind to ribosome and block elongation
Streptomycin inhibits peptide initiation and
elongation
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Mutations: Changes in the Genetic Code
A change in phenotype due to a change in
genotype (nitrogen base sequence of DNA) is
called a mutation.
A natural, nonmutated characteristic is known
as a wild type (wild strain).
An organism that has a mutation is a mutant
strain, showing variance in morphology,
nutritional characteristics, genetic control
mechanisms, resistance to chemicals, etc.
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Causes of Mutations
Spontaneous mutations random change
in the DNA due to errors in replication that
occur without known cause
Induced mutations result from exposure
to known mutagens, physical (primarily
radiation) or chemical agents that interact
with DNA in a disruptive manner
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Categories of Mutations
Point mutation addition, deletion or
substitution of a few bases
Missense mutation causes change in a
single amino acid
Nonsense mutation changes a normal
codon into a stop codon
Silent mutation alters a base but does not
change the amino acid

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Categories of Mutations
Back-mutation when a mutated gene
reverses to its original base composition
Frameshift mutation when the reading
frame of the mRNA is altered by the
addition or deletion of nucleotides in a
newly synthesized DNA
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Repair of Mutations
Since mutations can be potentially fatal, the cell
has several enzymatic repair mechanisms in
place to find and repair damaged DNA.
DNA polymerase proofreads nucleotides during
DNA replication
Mismatch repair locates and repairs mismatched
nitrogen bases that were not repaired by DNA
polymerase
Light repair for UV light damage
Excision repair locates and repairs incorrect
sequence by removing a segment of the DNA and
then adding the correct nucleotides
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The Ames Test
Any compound known to be mutagenic is
considered to be carcinogenic.
Agricultural, industrial, and medicinal
compounds are screened using the Ames test.
Indicator organism is a mutant strain of
Salmonella typhimurium that has lost the ability
to synthesize histidine.
This mutation is highly susceptible to back-
mutation.
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Positive and Negative Effects Of
Mutations
Mutations leading to nonfunctional proteins are
harmful, possibly fatal.
Organisms with mutations that are beneficial in
their environment can readily adapt, survive, and
reproduce these mutations are the basis of
change in populations.
Any change that confers an advantage during
selection pressure will be retained by the
population.

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DNA Recombination Events
Genetic recombination occurs when an
organism acquires and expresses genes
that originated in another organism
3 means for genetic recombination in bacteria:
1. Conjugation
2. Transformation
3. Transduction
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Conjugation
Conjugation transfer of a plasmid or
chromosomal fragment from a donor cell to
a recipient cell via a direct connection
Gram-negative cell donor has a fertility
plasmid (F plasmid, F factor) that allows the
synthesis of a conjugation (sex) pilus
recipient cell is a related species or genus
without a fertility plasmid
donor transfers fertility plasmid to recipient
through pilus
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Conjugation
High-frequency recombination donors
fertility plasmid has been integrated into the
bacterial chromosome
When conjugation occurs, a portion of the
chromosome and a portion of the fertility
plasmid are transferred to the recipient.
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Transformation
Transformation chromosome fragments
from a lysed cell are accepted by a recipient
cell; the genetic code of the DNA fragment is
acquired by the recipient
Donor and recipient cells can be unrelated
Useful tool in recombinant DNA technology
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Insert figure 9.23
transformation
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Transduction
Transduction bacteriophage serves as a
carrier of DNA from a donor cell to a recipient
cell
Two types:
generalized transduction random fragments of
disintegrating host DNA are picked up by the phage
during assembly; any gene can be transmitted this
way
specialized transduction a highly specific part of
the host genome is regularly incorporated into the
virus

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Transposons
Special DNA segments that have the
capability of moving from one location in
the genome to another jumping genes
Cause rearrangement of the genetic material
Can move from one chromosome site to
another, from a chromosome to a plasmid,
or from a plasmid to a chromosome
May be beneficial or harmful
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