Applications of transposon-based gene delivery system in bacteria.

ABSTRACT

Mobile genetic segments, or transposons, are also referred to as "jumping genes" as they can shift from one position in the genome to another, thus inducing a chromosomal mutation. According to the target site-specificity of the transposon during a transposition event, the result is either the insertion of a gene of interest at a specific chromosomal site, or the creation of knockout mutants. The former situation includes the integration of conjugative transposons via site-specific recombination, several transposons preferring a target site of a conserved ATrich sequence, and Tn7 being site-specifically inserted at attTn7, the downstream of the essential glmS gene. The latter situation is exploited for random mutagenesis in many prokaryotes, including IS (insertion sequence) elements, mariner, Mu, Tn3 derivatives (Tn4430 and Tn917), Tn5, modified Tn7, Tn10, Tn552, and Ty1, enabling a variety of genetic manipulations. Randomly inserted transposons have been previously employed for a variety of applications such as fmgenetic footprinting, gene transcriptional and translational fusion, signature-tagged mutagenesis(STM),DNA or cDNA sequencing, transposon site hybridization(TraSH), and scanning linker mutagenesis(SLM). Therefore, transposon-mediated genetic engineering is a baluable discipiline for the study of bacterial physiology and pathogenesis in living hosts.