BACTERIAL GENETICS
Genetics – study of heredity
Genotype – complete collection of genes
Phenotype – all the organism’s physical traits, attributes or
characteristics; presence or absence of enzymes and structures such
as capsules, flagella and pili
For example, an organism cannot produce a particular enzyme unless
it possesses the gene that codes for that enzyme. It cannot produce
flagella unless it possesses the gene necessary for flagella production.
Phenotype is the manifestation of the genotype
Genes direct all functions of the cell
The information in a gene is used by the cell to make an mRNA molecule (via the
process known as transcription). Then, the information in the mRNA molecule is used
to make a gene product (via the process known as translation).
Most gene products are proteins.
When the information in a gene has been used by the cell to make a gene product, the
gene coding for that particular gene product is said to have been expressed.
All the genes on the chromosome are not being expressed at any given time. It would
be pointless for a cell to produce a particular enzyme if that enzyme was not needed.
Genes that are expressed at all times are called constitutive genes.
Those that are expressed only when the gene products are needed are called inducible
genes.
The DNA of any gene on the chromosome is subject to accidental
alteration (e.g., the deletion of a base pair), which alters the gene
product and perhaps also alters the trait that is controlled by that
gene.
If the change in the gene alters or eliminates a trait in such a way
that the cell does not die or become incapable of division, the
altered trait is transmitted to the daughter cells of each
succeeding generation.
A change in the characteristics of a cell caused by a change in the
DNA molecule (genetic alteration) that is transmissible to the
offspring is called a mutation.
There are three categories of mutations:
A. beneficial mutations - of benefit to an organism,
B. harmful (and sometimes lethal) mutations - result in the
production of nonfunctional enzymes. Some harmful mutations are lethal
to the organism
C. silent mutation - causes no change in function of the cell,
have no effect on the cell
The mutation rate can be increased by exposing cells to physical or
chemical agents that affect the chromosome. Such agents are called
mutagens.
Spontaneous mutation
Random mutations that occur naturally
Mutation rate can be increased by exposing cells in the
physical or chemical agents that affect the chromosome
called mutagens
Mutant – organism containing the mutation
Mutagenic changes
May involve changes in cell shape, biochemical activities,
nutritional needs, antigenic sites, colony characteristics,
virulence and drug resistance
Ames test – a procedure which uses a strain of Salmonella
to learn whether a particular chemical is a mutagen
A mutagen may also be a carcinogen
In a test procedure called the Ames test (developed by Bruce Ames in the
1960s), a mutant strain of Salmonella is used to learn whether a particular
chemical (e.g., a food additive or a chemical used in some type of cosmetic
product) is a mutagen.
If exposure to the chemical causes a reversal of the organism’s mutation
(known as a back mutation), then the chemical has been shown to be
mutagenic.
If the chemical is mutagenic, then it might also be carcinogenic (cancer-
causing) and should be tested using laboratory animals or cell cultures.
Many substances found to be mutagenic by the Ames test have been shown to
be carcinogenic in laboratory animals.
Substances that are carcinogenic in laboratory animals might also be
carcinogenic in humans.
Ways in Which Bacteria Acquire New Genetic Information
1. Mutations (involve changes in the base sequences of genes)
2. Lysogenic conversion (involves bacteriophages and the acquisition of new viral genes)
3. Transduction (involves bacteriophages and the acquisition of new bacterial genes)
4. Transformation (involves the uptake of “naked” DNA)
5. Conjugation (involves the transfer of genetic information from one cell to another
involving what is known as a sex pilus)
Ways in which bacteria acquire new
genetic information
A. Mutation
B. Lysogenic conversion – involves a bacteriophage
Types of bacteriophage:
1. Virulent – causes lytic cycle
2. Temperate phage (lysogenic phage) – inject their DNA
into the bacterial cell, the phage DNA integrates into the
bacterial chromosome but does not cause lysis
After temperate phages (also known as lysogenic phages) inject their DNA into the
bacterial cell, the phage DNA integrates into (becomes part of) the bacterial
chromosome but does not cause the lytic cycle to occur. This situation— in which the
phage genome is present in the cell but is not causing the lytic cycle to occur—is
known as lysogeny.
During lysogeny, all that remains of the phage is its DNA; in this form, the phage is
referred to as a prophage.
The bacterial cell containing the prophage is referred to as a lysogenic cell or lysogenic
bacterium.
Each time a lysogenic cell undergoes binary fission, the phage DNA is replicated along
with the bacterial DNA and is passed on to each of the daughter cells. The daughter
cells are also lysogenic cells.
Lysogenic conversion
Although the prophage does not usually cause the lytic cycle to occur, certain events (e.g.,
exposure of the bacterial cell to ultraviolet light or certain chemicals) can trigger it to do
so.
While the prophage is integrated into the bacterial chromosome, the bacterial cell can
produce gene products that are coded for by the prophage genes.
The bacterial cell will exhibit new properties—a phenomenon known as lysogenic
conversion (or phage conversion).
In other words, the bacterial cell has been converted as a result of lysogeny and is now
able to produce one or more gene products that it previously was unable to produce.
A lysogenic bacterium is capable of producing one or more new gene products
as a result of infection by a temperate bacteriophage
Examples: Corynebacterium diphtheriae – causes diphtheria
Streptococcus pyogenes – causes scarlet fever
Clostridium botulinum – causes food poisoning
Vibrio cholerae – causes cholera
Ways in which bacteria acquire new
genetic information
C. Transduction – means carry across; also involves
bacteriophage
Some bacterial genetic material may be carried across from
one bacterial cell to another by a bacterial virus. This
phenomenon may occur after infection of a bacterial cell by
a temperate bacteriophage
The viral DNA of the temperate bacteriophage combines
with the bacterial DNA, thus becoming a prophage
If a stimulating chemical, heat, or ultraviolet light activates the prophage, it begins to
produce new viruses via the production of phage DNA and proteins.
As the chromosome disintegrates, small pieces of bacterial DNA may remain attached
to the maturing phage DNA.
During the assembly of the virus particles, one or more bacterial genes may be
incorporated into some of the mature bacteriophages.
When all the phages are released by cell lysis, they proceed to infect other cells, some
injecting bacterial genes as well as viral genes.
Bacterial genes that are attached to the phage DNA are carried to new cells by the
virus.
Ways in which bacteria acquire new
genetic information
D.Transformation – a bacterial cell becomes
genetically transformed after the uptake of naked
DNA fragments
Itwas first demonstrated by Frederick Griffith ad
his colleagues in 1928
In 1944, Oswald Avery and his colleagues proved
that DNA was the molecule that contained genetic
information
Oswald Avery Experiment on
Transformation
DNA extract from encapsulated, pathogenic Streptococcus pneumoniae bacteria (referred
to as S. pneumoniae type 1) was added to nonencapsulated, nonpathogenic S.
pneumoniae (referred to as S. pneumoniae type 2).
At the beginning of the experiment, there were no live encapsulated bacteria in the
culture. After incubation, live type 1a broth culture of encapsulated) bacteria were
recovered from the culture.
The only possible explanation was that some of the live type 2 bacteria must have taken
up (absorbed) some of the type 1 DNA from the broth.
Type 2 bacteria that absorbed pieces of type 1 DNA containing the gene(s) for capsule
production were now able to produce capsules.
Type 2 (nonencapsulated) bacteria were converted to type 1 (encapsulated) bacteria as a
result of the uptake of the genes that code for capsule production.
Transformation can occur in Bacillus, Escherichia,
Haemophilus, Pseudomonas and Neisseria
It may also occur between two species (Example: between
Staphylococcus and Streptococcus)
The ability to absorb naked DNA into the cell is referred to
as competence, and bacteria capable of taking up naked
DNA molecules are said to be competent bacteria.
Conjugation
Discovered by Joshua Lederberg and Edward Tatum in 1946 while
experimenting on E. coli
Conjugation involves a specialized type of pilus called a sex pilus
(sometimes referred to as an F pilus or a conjugation bridge).
A bacterial cell (called the donor cell or F+ cell) possessing a sex pilus
attaches to another bacterial cell (called the recipient cell or F− cell)
by means of the sex pilus
The genetic material in the form of a plasmid is transferred through
the hollow sex pilus from a donor to a recipient cell.
A plasmid that contains multiple genes for antibiotic resistance is
called a resistance factor or R-factor
Conjugation occurs mostly among species of enteric, gram negative
bacilli and in species of Pseudomonas and Streptococcus
Pilus retraction brings the two cells into tight contact. Within the
donor cell, a enzyme called relaxase nicks the double-stranded F
plasmid DNA and guides one of the strands to a coupling protein
the DNA pump).
The single DNA strand is then transferred from the donor cell to
the recipient cell through a conjugative pore, which forms at the
juncture of the two bacterial cells
Conjugation occurs mostly among species of enteric, Gram-negative bacilli,
but has also been reported within species of Pseudomonas and Streptococcus.
Sex pili are thicker and longer than other pili.
Genes that may be transferred by conjugation, include those coding for
antibiotic resistance, colicin (a protein produced by E. coli that kills certain
other bacteria), and fertility factors (F+ and Hfr+), where F stands for fertility
and Hfr stands for high frequency of recombination.
If a plasmid contains multiple genes for antibiotic resistance, the plasmid is
referred to as a resistance factor or R-factor.
A recipient cell that receives an R-factor becomes a multiply drug-resistant
organism (referred to by the press as a “superbug”).
Genetic engineering (Recombinant DNA
technology)
An array of techniques to transfer genes into other easily
cultured cells to facilitate the large-scale production of
important gene products
Plasmids – vectors used for inserting genes into cells
Examples: insulin, somatostatin (inhibits the release of
the growth hormone called somatotropin); interferon,
plasminogen-activating factor
Genetically engineered microorganisms can also be used
to clean up the environment (e.g., to get rid of toxic
wastes).
Examples in Genetic Engineering
A soil bacterium contains a gene that enables the organism to
break oil down into harmless by-products, but, because the
organism cannot survive in salt water, it cannot be used to clean
up oil spills at sea. Remove the gene from the soil bacterium, and,
using a plasmid vector, insert it into a marine bacterium. Now the
marine bacterium has the ability to break down oil and, in large
numbers, can be used to clean up oil spills at sea.
In medicine, there is potential for making engineered antibodies,
antibiotics, and drugs; for synthesizing important enzymes and
hormones for treatment of inherited diseases; and for making
vaccines. Such vaccines would contain only part of the pathogen
(e.g., the capsid proteins of a virus) to which the person would
form protective antibodies
Gene Therapy
Insertion of a normal gene into cells to correct a specific
genetic or acquired disorder caused by a defective gene
The first gene therapy trials were conducted in the US in
1990
Viral delivery is the most common method for inserting
genes into cells
Viruses are used as vectors. Examples: adeno-associated
viruses; retroviruses; adenoviruses and herpesviruses
