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Fig. 7.1
Plasmids rely on the DNA- replication enzymes of the host cell for their reproduction,
but the initiation of replication is controlled by plasmid genes.
The pilus between the E. coli cells above is an F pilus whose synthesis results from
the presence of a conjugative plasmid called the F factor (F stands for fertility).
Cells that contain the F plasmid (low-copy number plasmid) are donors and are
designated the F+ cells (F plus); those lacking F are recipients and are designated
the F- cells (F minus.)
Conjugation begins with physical contact between a donor cell and a recipient cell.
Once the pilus contacts the F- cell, the pilus retracts and the cell membranes of the
donor and recipient are brought into close proximity. Then the donor DNA moves
through a pore in the membrane from the donor to the recipient. The transfer is
always accompanied by replication of the plasmid. Contact between an F+ and an
F- cell initiates rolling circle replication which results in the transfer of a singlestranded linear branch of the rolling circle to the recipient cell. During transfer, DNA
is synthesized in both donor and recipient. When transfer is complete, the linear F
strand becomes circular again in the recipient cell. After the transfer, both cells
contain F and can function as donors. The F- cell has been converted into an F+ cell.
Fig. 7.3
Transposable elements are DNA sequences that can jump from one position to
another or from one DNA molecule to another. Bacteria contain a wide variety of
transposable elements. The smallest and simplest are insertion sequences or IS
elements, which are typically 1-3 kb in length and usually encode only the
transposase protein required for transposition and one or more additional proteins
that regulate the rate of transposition.
Other transposable elements in bacteria contain one or more genes unrelated to
transposition that can be mobilized along with the transposable element; this type
of element is called a transposon. Much of the widespread antibiotic resistance
among bacteria is due to the spread of transposons that include one or more
antibiotic- resistance genes. When a transposon mobilizes and inserts into a
conjugative plasmid, it can be widely disseminated among different bacterial hosts
by means of conjugation.
Some transposons have composite structures with antibiotic resistance sandwiched
between insertion sequences, as is the case with the Tn5 element (illustration
below), which terminates in two IS 50 elements in inverted orientation. Transposons
are designated by the abbreviation Tn followed by an italicized number (ex. Tn5).
For example, Tn5( neo- r ble-r str-r) contains genes for resistance to three different
antibiotics: Neomycin, Bleomycin and Streptomycin.
Fig. 7.4
Nonconjugative and conjugative plasmids typically coexist in the same cell along
with host genomic DNA, and when a transposable element is mobilized, all of the
DNA molecules present are potential targets for insertion. Many nonconjugative and
conjugative plasmids present in a bacterial cell come to carry one or more copies of
the same transposable element. Because these copies are homologous DNA
sequences, they can serve as substrates for recombination. When 2 plasmids
undergo recombination in a region of homology, the result is as shown below. The
recombination forms a composite plasmid called a cointegrate. By this
mechanism, nonconjugative plasmids can temporarily ride along with conjugative
plasmids and be transferred from cell to cell.
Fig. 7.5
In the evolution of multiple antibiotic resistance, bacteria have also made liberal use
of a set of enzymes known as Site- specific recombinases which were present in
bacterial populations and functioned in the evolution of other traits long before the
antibiotic era. Each type of site specific recombinase binds with a specific
nucleotide sequence in duplex DNA. When the site is present in each of two duplex
DNA molecules, the recombinase brings the sites together and catalyzes a
reciprocal exchange between the duplexes. Site specific recombinases are used in
the assembly of multiple antibiotic-resistance units called integrons. An integron
is a DNA element that encodes a site- specific recombinase as well as a recognition
region that allows other sequences with similar recognition regions to be
incorporated into the integron by recombination. The elements that integrons
acquire are known as cassettes. In the