Bacterial Recombination

Bacterial Recombination
• Genetic recombination refers to the exchange of genes between two
DNA molecules to form new combinations of genes on a
chromosome.

• Like mutation, genetic recombination contributes to a population„s

genetic diversity, which is the source of variation in evolution.

• In highly evolved organisms such as present-day microbes,

recombination is more likely than mutation to be beneficial because
recombination will less likely destroy a gene's function and may
bring together combinations of genes that enable the organism to
carry out a valuable new function.
 Bacterial Recombination
• Vertical gene transfer – From parents to offspring.

• Horizontal gene transfer – From one microbe to another.

• Part of total DNA from Donor cell integrated into Recipient cell.

• Remaining amount of DNA from donor cell degraded.

• Recipient cell with DNA from donor is called Recombinant.

• 1% of population might undergo Recombination.

 Bacterial Recombination
• Transformation.

• Conjugation.

• Transduction.
 Transformation
• Transfer of naked DNA from donor to recipient cell.

• Transformation experiment by Griffith showed that DNA is the

genetic material and can be transferred between host and recipient
DNA.

• E.coli cannot undergo transformation naturally, hence it is made

competent in the lab.

• The process is called „Artificial Transformation‟.

 Transformation
• Bacterial transformation done without mice.

• Broth containing non-encapsulated living bacteria, to which

dead encapsulated bacteria were added.

• After incubation, encapsulated living virulent bacteria were found.

• This proves that non-encapsulated bacteria received genes

from dead encapsulated ones and got genes for forming a capsule.
 Transformation
• The material responsible for transmission of this character was not
known.

• In 1944, Oswald T Avery, Colin M Macleod, Maclyn Mccarty

proved that DNA is the genetic material.

Peter J. Russell, iGenetics: Copyright © Pearson Education, Inc., publishing as Benjamin Cummings.
Transformatio
n
 Transformation
• After death, cell lysis leads to release of DNA from bacteria.

• Other bacteria take up DNA and integrate into their chromosomes

by recombination.

• recA protein binds to donor and cells DNA and causes exchange of
strands.

• Recipient cell with this combination of genes will now become a

hybrid or recombinant.

• All its daughter cells will be recombinant.

 Transformation
• Bacillus, Haemophilus, Streptococcus, Staphylococcus, Neisseria
etc. undergo transformation in nature.

• Transformation works best when both donor and recipient

are closely related.

• A small portion of DNA is transferred, which is still large to cross

the cell wall and membrane in the recipient cell.
 Conjugation
• Needs extra chromosomal elements called Plasmids.

• Plasmids replicate independently of chromosome.

• They carry non-essential genes for growth during normal conditions.

• They give advantage for cells during stress.

• Ex: Antibiotic resistance genes.

• Plasmids can be transferred from one cell to another ( Conjugative

or transferable plasmids).
 Conjugation vs. Transformation
• Conjugations needs direct cell to cell contact.

• Conjugating cells must be of opposite mating type.

• Donor cells carry plasmids, recipient cells don‟t.

• Gram negative bacteria produce sex pili which contacts both cells
directly.

• Gram positive bacteria produce sticky surface molecules that bring

two cells in contact.
 Conjugation
• Single strand is transferred from donor to recipient.

• In the recipient the SS plasmid is replicated.

• In E.coli, Fertility (F) Factor was the first plasmid observed to be

transferred.

• Donor cells with F factor are F+ cells, recipients without F factor are
F- cells.

• Donor cells transfer F factor to recipient cell, hence recipient cells

become F+ cells.
 Conjugation
• In donor cells, F factor may integrate into the host
chromosome becoming hfr (High Frequency of Recombination).

• Thus F+ cells become hfr cells.

• Conjugation between hfr and F- cells results in replication of the

chromosome with F factor.

• A single parental strand is transferred from hfr cell to the F- cells.

 Conjugation
• Complete transfer of the chromosome does not take place.

• Only a small piece of F factor leads the chromosomal genes into F-

cells.

• The small strand containing chromosomal genes recombines with

the DNA of F- cells.

• Thus F- cells receive only a part of chromosomal genes and hence do

not get converted to F+ cells.
 Transduction
• Transduction is the process of moving bacterial DNA from one cell
to another using a bacteriophage.

• Bacteriophage or just “phage” are bacterial viruses.

• They consist of a small piece of DNA inside a protein coat.

• The protein coat binds to the bacterial surface, then injects the phage
DNA.

• The phage DNA then takes over the cell‟s machinery and
replicates many virus particles.
Transductio
n
 Transduction
1. Phage attaches to the cell and injects its DNA.

2. Phage DNA replicates, and is transcribed into RNA,

then translated into new phage proteins.

3. New phage particles are assembled.

4. Cell is lysed, releasing about 200 new phage particles.

Total time = about 15 minutes.

 Generalized Transduction
• Some phages, such as phage P1, break up the bacterial chromosome
into small pieces, and then package it into some phage particles
instead of their own DNA.

• These chromosomal pieces are quite small.

• A phage containing E. coli DNA can infect a fresh host, because the
binding to the cell surface and injection of DNA is caused by the
phage proteins.
 Generalized Transduction
• After infection by such a phage, the cell contains an exogenote
(linear DNA injected by the phage) and an endogenote (circular
DNA that is the host‟s chromosome).

• A double crossover event puts the exogenote‟s genes onto the

chromosome, allowing them to be propagated.
 Specialized Transduction
• Some phages can transfer only particular genes to other bacteria.

• Phage lambda (λ) has this property. To understand specialized

transduction, we need to examine the phage lambda life cycle.

• lambda has 2 distinct phases of its life cycle. The “lytic” phase : the
phage infects the cell, makes more copies of itself, then lyses the
cell to release the new phage.
 Specialized Transduction
• The “lysogenic” phase of the lambda life cycle starts the same way:
the lambda phage binds to the bacterial cell and injects its DNA.

• Once inside the cell, the lambda DNA circularizes, then incorporates
into the bacterial chromosome by a crossover.

• Once incorporated into the chromosome, the lambda DNA becomes

quiescent: its genes are not expressed and it remains a passive
element on the chromosome, being replicated along with the rest of
the chromosome.

• The lambda DNA in this condition is called the “prophage”.

 Specialized Transduction
• After many generations of the cell, conditions might get harsh.
For lambda, bad conditions are signaled when DNA damage
occurs.

• When the lambda prophage receives the DNA damage signal, it

loops out and has a crossover, removing itself from the
chromosome. Then the lambda genes become active and it goes
into the lytic phase, reproducing itself, then lysing the cell.
Lysogenic life cycle
 Specialized Transduction
• lambda can only incorporate into a specific site, called
attachment site attλ. The Phase ATT site and bacterial ATT sites are
similar and can complex with each other.

• The gal gene is on one side of attλ and the bio gene (biotin
synthesis) is on the other side.

• When happens, a piece of the E. coli chromosome is

incorporated
this into the lambda phage chromosome
• Sometimes when lambda come out of the chromosome at the end of
the lysogenic phase, it crosses over at the wrong point.
 Specialized Transduction
• These phage that carry an E. coli gene in addition to the lambda
genes are called “specialized transducing phages”.

• They can carry either the gal gene or the bio gene to other E. coli.