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Triparental mating is a form of
Protocol for P. aeruginosa
bacterial conjugation
Bacterial conjugation is the transfer of genetic material between Bacteria, bacterial cells by direct cell-to-cell contact or by a bridge-like connection between two cells. This takes place through a pilus. It is a parasexual cycle, parasexual mode ...
where a conjugative plasmid present in one bacterial strain assists the transfer of a mobilizable plasmid present in a second bacterial strain into a third bacterial strain. Plasmid
A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. They are most commonly found as small circular, double-stranded DNA molecules in bacteria and ...
s are introduced into bacteria for such purposes as transformation, cloning
Cloning is the process of producing individual organisms with identical genomes, either by natural or artificial means. In nature, some organisms produce clones through asexual reproduction; this reproduction of an organism by itself without ...
, or transposon
A transposable element (TE), also transposon, or jumping gene, is a type of mobile genetic element, a nucleic acid sequence in DNA that can change its position within a genome.
The discovery of mobile genetic elements earned Barbara McClinto ...
mutagenesis. Triparental matings can help overcome some of the barriers to efficient plasmid mobilization. For instance, if the conjugative plasmid and the mobilizable plasmid are members of the same incompatibility group they do not need to stably coexist in the second bacterial strain for the mobilizable plasmid to be transferred.
History
Triparental mating was identified in yeasts in 1960 and then in ''Escherichia coli
''Escherichia coli'' ( )Wells, J. C. (2000) Longman Pronunciation Dictionary. Harlow ngland Pearson Education Ltd. is a gram-negative, facultative anaerobic, rod-shaped, coliform bacterium of the genus '' Escherichia'' that is commonly fo ...
'' in 1962.
Process
Requirements
*A ''helper strain'', carrying a conjugative plasmid (such as the F-plasmid) that codes for genes required forconjugation
Conjugation or conjugate may refer to:
Linguistics
*Grammatical conjugation, the modification of a verb from its basic form
*Emotive conjugation or Russell's conjugation, the use of loaded language
Mathematics
*Complex conjugation, the change o ...
and DNA
Deoxyribonucleic acid (; DNA) is a polymer composed of two polynucleotide chains that coil around each other to form a double helix. The polymer carries genetic instructions for the development, functioning, growth and reproduction of al ...
transfer.
*A ''donor strain'', carrying a mobilizable plasmid that can utilize the transfer functions of the conjugative plasmid.
*A ''recipient strain'', you wish to introduce the mobilizable plasmid into.
Five to seven days are required to determine if the
plasmid was successfully introduced into the new bacterial
strain and confirm that there is no carryover of the helper or
donor strain.
In contrast, electroporation
Electroporation, also known as electropermeabilization, is a microbiological and biotechnological technique in which an electric field is applied to cells to briefly increase the permeability of the cell membrane. The application of a high-vo ...
does not require a helper or
donor strain. This helps
avoid possible contamination with other strains. The introduction
of the plasmid can be verified in the recipient
strain in two days, making electroporation a faster and
more efficient method of transformation. Electroporation however does not work with all bacteria and is mostly limited to well-characterized model organisms.
See also
*Bacterial conjugation
Bacterial conjugation is the transfer of genetic material between Bacteria, bacterial cells by direct cell-to-cell contact or by a bridge-like connection between two cells. This takes place through a pilus. It is a parasexual cycle, parasexual mode ...
* Plasmid
A plasmid is a small, extrachromosomal DNA molecule within a cell that is physically separated from chromosomal DNA and can replicate independently. They are most commonly found as small circular, double-stranded DNA molecules in bacteria and ...
* Transposon (applications)
* Bacteriophage
A bacteriophage (), also known informally as a phage (), is a virus that infects and replicates within bacteria. The term is derived . Bacteriophages are composed of proteins that Capsid, encapsulate a DNA or RNA genome, and may have structu ...
* Three-parent baby
Mitochondrial replacement therapy (MRT), sometimes called mitochondrial donation, is the replacement of Mitochondrion, mitochondria in one or more cells to prevent or ameliorate disease. MRT originated as a special form of in vitro fertilisation ...
External links
Protocol for P. aeruginosa
References
{{Reflist Molecular biology