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Transduction

Transduction is a process of transfer of bacterial genes through bacteriophages.


Discovery
In 1951, Joshua Lederberg and Norton Zinder were repeating the experiments of
Lederberg and Tatum (1946), i.e. conjugation experiment in E.coli k12. They were
working on Salmonella typhimurium using the same technique-U arm tube with
membrane filter. The intent of their study was to confirm conjugation in Salmonella.
However, they happen to discover transduction in bacteria, a new means of
recombination in bacteria.

Lederberg & Zinder experiment


• Transduction was first discovered in 1952 by Joshua Lederberg and Norton
Zinder
– Mixed two strains of bacteria
• LA22 strain: phe- trp- met+ his+
• LA2 strain :phe+ trp+ met- his-
– Plated mixture on minimal media
• Media lacked these four amino acids
– 1 cell in 100,000 grew
• Genotype phe+ trp+ met+ his+
• Genetic material had been transferred
The researchers have found that when two strains were mixed, prototrophs were obtained
at a frequency 10-5. The prototrophs could synthesize all amino acids
(phe+trp+met+his+). This demonstrated genetic recombination in S. typhimurium
conjugation.
Confirmation that recombination was not through conjugation but through other
mode, transduction
In order to confirm that recombination occurred via process other than conjugation, the
researchers have prevented cell contact by using a filer between the two arm.
Each strain was added in U arm tube. The two arms of the U tube were separated
by a bacteria proof sintered glass filter, which will prevent contact between two bacteria
but will allow free movement of medium. The suction was applied so that culture
medium can pass from one arm to other. Thereafter, fluid was drawn from both arm and
plated on M.M. media. The prototrophs were recovered from fluid drawn from the arm
where LA22 culture was placed but not from the fluid drawn from arm where LA2
culture was placed. From LA2 culture, a filterable agent was isolated which was resistant
to DNase. This later was identified as P22 virus. The filterable agent was sensitive to
antiserum of P22. In the presence of antiserum, no prototrophs were recovered from
strain LA22. This confirms the involvement of virus mediated gene transfer from one
strain of bacteria to the other strain of bacteria. Transduction is usually intragenic-
involving closely related strains of bacteria but intergeneric transduction have also been
reported, i.e. E. coli and Salmonella, E. coli and Shigella.
Types of Ttransduction
1. Generalized Transduction
2. Specialized Transduction

1. Generalized Transduction: Transduction in which potentially any donor bacterial


gene can be transferred. It is shown by virulent phages or lysogenic phages during its
lytic conversion where it picks a DNA fragment from a bacterium transfer it to
another bacterium during infection by the phage. During packaging instead of phage
DNA to be packaged, fragment of bacterial DNA is packaged inside phage head. P22
phages can package 1% of bacterial chromosome. The extent of packaging of
bacterial DNA depends upon the phage head. If it is of larger size, it can
accommodate more bacterial genome. If the phage head is small only small portion of
bacterial chromosome can be packaged. The frequency of transduction ranged from
10-5 to 10-7.
Steps in Generalized Transduction

1. Infection of Donor:

2. Phage replication and degradation of host DNA


3. Assembly of phages particles: During a LYTIC infection, a transducing phage,
such as P1 infecting E. coli, accidentally packages a piece of the bacterial
chromosome into a virus particle instead of its own viral DNA.

4. Release of phage
5. Infection of recipient: The phage carrying the bacterial DNA then delivers it to the
recipient cell when it tried to infect again
6.
7. Legitimate recombination: The injected bacterial DNA may then be inserted into
recipient chromosome by homologous recombination
Fig. Complete
Transduction by
transducing phage P1
Abortive Transduction: In abortive transduction, a segment of bacterial DNA
(exogenote) is introduced into another bacterium by a bacteriophage. When the
exogenote is integrated into the recipient bacteria, this is called “complete
transduction”. But when the exogenote is not integrated into endogenote and remains
free, it is called “ abortive transduction”. When the abortive transductant divide, out of
the two-only one daughter cells contain exogenote. The other cell do not contain
exogenote. As the time progresses, the exogenote is progressively diluted out through
subsequent cell divisions. This type of transduction is known as ‘abortive
transduction”.
Normal Events Leading to Lysogeny
The steps leading to lysogeny under normal circumstances follows as:
• Circularization of the phage chr
– Cohesive ends

•Site-specific recombin
Cohesive Ends
ly

• Site-specific
recombination
Linear Double Stranded Opened C
– Phage coded
enzyme
gal
– λ phage
integrates by
Termination
• Induction
– Adverse conditions
• Role of proteases g

– recA protein
– Destruction of
repressor
• Gene expression
• Excision
Specialized Transduction- error in lysogenic cycle.

• Error in L

• Excision of the
prophage
• Replication and
release of phage
• Infection of the
During specialized transduction, only specific gene from donor bacterium is transferred
to the recipient bacterium through phages. Unlike random packaging of bacterial DNA,
this form of gene exchange results from imprecise excision of an integrated phage
(prophage).

λ phage integra
recombination (

gal B
λ bio or λ gal trans
imprec

gal B P’

λ
bio
B’ P
Due to imprecise excision, lemda bio or lemda gal transducing
phage Phages that carry a very small part of the adjacent flanking bacterial
genome, either to the left or right of the att site. These phage can
recombine into the genome via phage –mediated integration or
homologous recombination.