Gene Transfer In Bacteria-1

Conjugation
 (F+ x F- conjugation, Hfr Conjugation, 
F’ Conjugation or Sexduction)
Mechanism of genetic exchange in bacteria

Bacteria exchange
genetic material
through three
different parasexual
processes
Conjugation
 Conjugation in bacteria was discovered in 1946 by J Lederberg and E.L.
Tatum and confirmed by Bernard Davis in 1950 by the famous U-tube
experiment which showed that conjugation required physical contact.
Conjugation
Conjugation is a process which
involves physical matting between
sexually differentiated strains of
Donor
bacteria for the unidirectional
Recipient F+ cell
transfer of genetic material.
F- cell
Or
Conjugation is a one- way transfer
of genetic material in bacteria from
a donor (male cell) to recipient
(female cell) by formation of close
physical contact (Pilus).
It is parasexual reproduction
(parasexual because it does not
involve formation of true gametes or
gamete fusion) in bacteria.
Properties

For conjugation close physical contact between bacterial cells are needed.

Gene transfer is unidirectional from donor (F+ cell) to recipient (F-cell).

Donor F+ cells carry a plasmid called the fertility factor or F factor or F plasmid a
circular extrachromosomal molecule containing about 94,000 np.

Genes on the F factor are responsible for facilitating cell-cell contact and F factor transfer
from donor to recipient cell.

Many F factor genes direct the synthesis of sex pili (singular pilus), a filamentous
appendages on the bacterial surface that attach F+­ cells to F- cells.

The F factor and the main bacterial chromosome have insertion sequences (ISs) that
enable the F factor to insert into the chromosome by homologous recombination.

Insertion sequences are short, transposable DNA sequences that contain a gene encoding
an enzyme called transposase which the movement of the insertion sequence around the
genome.
Types of Conjugation

 F+ x F- Conjugation
 Hfr Conjugation
 F’ Conjugation of Sexduction
 F+ x F- Conjugation
 During F+ x F- Conjugation, the F factor directs synthesis of transfer
proteins which are encoded by tra region.
 Tra proteins are needed for synthesis of pilus and replication of F factor by
rolling circular mechanism using OriT as origin of replication sequences.
 The F pili of a donor cell make contact with a recipient cell that lacks an F
factor and attach to that cell, so that the two cells can be pulled into close
contact.
 A conjugation tube forms between the cells.
 The F factor started replicating at OriT by rolling circle mechanism of
replication.
 The newly synthesized single stranded DNA is transferred to F- cell via
conjugation tube. The single stranded circular DNA in F- cell converted to
double stranded circular F factor making F-cell to F+ cell.
 So the result of F+ x F- Conjugation is recipient F- cell become F+ cell.
 If a population of F+ cells is mixed with a population of F- cells, virtually
all of the cells will acquire an F factor.
The diagrammatic representation of F+ x F- Conjugation:
Hfr Conjugation
 The F factor can exist in either of two states:
 Autonomous state, in which it replicates independently of the bacterial
chromosome.
 Integrated state, in which it is covalently inserted into the bacterial chromosome
and replicates like any other segment of that chromosome.
 The F factor can integrate into the bacterial chromosome by site-specific
recombination events.
 The integration of the F factor is mediated by short DNA sequences (insertion
sequences) that are present in multiple copies in both the F factor and the bacterial
chromosome.
 Thus, an F factor can integrate at many different sites in the bacterial chromosome.
 A cell that carries an integrated F factor is called an Hfr cell (for high-frequency
recombination).
 In its integrated state, the F factor mediates the transfer of the chromosome from
the Hfr cell to a recipient (F-) cell during conjugation.
 Usually, the cells separate before chromosome transfer is complete; thus, only
rarely will an entire chromosome be transferred from an Hfr cell to a recipient cell.
The formation of an Hfr cell by the
integration of an autonomous F factor.
The F factor is covalently inserted into
the chromosome by site-specific
recombination between homologous
DNA sequences in the F factor and the
chromosome
Transfer of bacterial genes from a Hfr cell to a
F– cell in Hfr x F- conjugation
Hfr x F- conjugation
 The mechanism that transfers DNA from a Hfr cell to a recipient cell
during conjugation appears to be the same if just the F factor is being
transferred, as in F+ to F- matings,
 The DNA transfer is initiated at the origin of replication (OriT) on
the F factor.
 Rolling circular replication transfer the donor strand from the main
chromosome into recipient cell where it become double stranded.
 Usually only a small part of the F factor is transferred to recipient,
and the recipient remains F- cell.
 It takes about 100 minutes for the entire bacterial chromosome to be
transferred, and the connection between conjugating cells is usually
severed before the process is finished
 Therefore the F- cells remain F- after conjugation process however
become partial diploid for many chromosomal genes.
F’ Conjugation or Sexduction
 The integrated F factor in an Hfr cell may exit the bacterial chromosomal by
reversing the steps that resulted in its integration.
 Sometime there is an error in the excision of the F factor from the
chromosome. This error result in the formation of an F’ factor, an F factor
containing a segment of the bacterial chromosome.
 The mating of an F’ cell with F- cell is basically the same as an F+ x F- mating,
with one important difference: bacterial genes incorporated in the F’ factor are
transferred with high frequency to recipient cell.
 These genes do not have to be integrated into recipient cell’s chromosome to
be expressed. The recipient acquires the F’ factor and become partially diploid
for the bacterial genes that it carries. The transfer of bacterial genes by F’
factor is called sexduction.
 Sexduction results in the rapid spread of bacterial genes from the main
chromosome through a population
Excision of F’ factor (F factor with bacterial gene(s)) from Hfr chromosome and
Sexduction (the transfer of bacterial genes by F’ factor)
References
 Principles of Genetics Sixth Edition by D. Peter
Snustad and  Michael J. Simmons, John Wiley &
Sons, Inc.
 Genetics: A conceptual Approach by Benjamin A.
Pierce, 3rd edition 2009, WH Freeman and Company.
 Microbiology Fifth Edition, 2002, by Prescott,
Harley, and Klein, The McGraw−Hill Companies