A gene cassette is a type of mobile genetic element that contains a gene and a recombination site. Since each cassette usually only contains a single gene these genetic elements tend to be very small; 500–1000 base pairs. They may exist incorporated into an integron or freely as circular DNA. Gene cassettes can move around within an organisms genome or be transferred to another organism in the environment via horizontal gene transfer. These cassettes often carry antibiotic resistance genes. An example would be the kanMX cassette which confers kanamycin (an antibiotic) resistance upon bacteria.
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Integrons
Integrons are genetic structures in bacteria which express and are capable of acquiring and exchanging gene cassettes. The integron consists of a promoter, an attachment site, and an integrase gene that encodes a site-specific recombinase The mobile units that insert into integrons are gene cassettes. For cassettes that carry a single gene without a promoter, the entire series of cassettes is transcribed from an adjacent promoter within the integron. The gene cassettes are speculated to be inserted and excised via a circular intermediate. This would involve recombination between short sequences found at their termini and known as 59 base elements (59-be)—which may not be 59 bases long. The 59-be are a diverse family of sequences that function as recognition sites for the site-specific integrase (enzyme responsible for integrating the gene cassette into an integron) that occur downstream from the gene coding sequence.
Genetic engineering
In genetic engineering, a gene cassette is a manipulable fragment of DNA carrying, and capable of expressing, one or more genes of interest between one or more sets of restriction sites. It can be transferred from one DNA sequence (usually on a vector) to another by 'cutting' the fragment out using restriction enzymes and 'pasting' it back into the new context. The vectors containing the gene of interest typically carry an antibiotic resistance gene also to easily identify cells that have successfully integrated the vector in to their genome.
To introduce a vector in to a target cell, a state of competence must be inferred on the cell. This state is induce in the lab by incubating cells with calcium chloride before a brief heat shock, or by electroporation. This makes the cells more susceptible to the plasmid that is being inserted. Once the plasmid has been added, the cells are grown in the presence of an antibiotic to confirm the uptake and expression of the new genetic elements.
Horizontal gene transfer
Horizontal gene transfer (HGT) is the transfer of genetic elements between cells other than parental inheritance. HGT is responsible for much of the spread of antibiotic resistance among bacteria. Gene cassettes containing antibiotic resistance genes, or other virulence factors such as exotoxins, can be transferred from cell to cell via phage, transduction, taken up from the environment, transformation, or by bacterial conjugation. The ability to transfer gene cassettes between organisms has played a large role in the evolution of prokaryotes.