首页|New Bacillus subtilis vector,pSSp,as genetic tool for site-specific integration and excision of cloned DNA,and prophage elimination
New Bacillus subtilis vector,pSSp,as genetic tool for site-specific integration and excision of cloned DNA,and prophage elimination
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NSTL
Microbiol Res Foundation
Site-specific recombination(SSR)systems are employed in many genetic mobile elements,including temperate phages,for their integration and excision.Recently,they have also been used as tools for applications in fields ranging from basic to synthetic biology.SPp is a temperate phage of the Siphoviridae family found in the laboratory standard Bacillus subtilis strain 168.SPβ encodes a ser-ine-type recombinase,SprA,and recombination directionality factor(RDF),SprB.SprA catalyzes recombination between the attachment site of the phage,attP,and that of the host,attB,to integrate phage genome into the attB site of the host genome and generate attL and attR at both ends of the prophage genome.SprB works in conjunction with SprA and switches from attBlattP to attL/R recombination,which leads to excision of the prophage.In the present study,we took advantage of this highly efficient recombination system to develop a site-specific integration and excision plasmid vector,named pSSp.It was constructed using pUC plasmid and the SSR system components,attP,sprA and sprB of SPp.pSSp was integrated into the attB site with a significantly high efficiency,and the resulting pSSp-integrated strain also easily eliminated pSSp itself from the host genome by the induction of SprB expression with xylose.This report presents two applications using pSSp that are particularly suitable for gene complementation experiments and for a curing system of SPp prophage,that may serve as a model system for the removal of prophages in other bacteria.
NSTL
Microbiol Res Foundation
