원문정보
초록
영어
Chromosomal gene inactivation was generally achieved via homologous recombination using heterologous or conditionally replicating vectors, or various site-specific recombination systems such as Flp-FRT, Gateway (Invitrogen), ParA-res , TnpR-res, and Cre-lox. But efficient gene
inactivation tools have not been developed in Leucnostoc(Ln.) yet.
In this study, a new site-specific insertional inactivation method was established in Ln. citreum CB2567 by using pLC4255 vector and it was used to delete dextransucrase gene (dsr) via homologous recombination.
Dextransucrase-inactivated mutant Ln. citreum CBM-D2567 was selected on agar plate with its non-dextran phenotype. PCR amplification of fragments using correspondence primers, quantification of dextran, cell scanning electromicroscope, dextransucrase activity analysis, and SDS-PAGE analysis of the mutant in sucrose medium indicated deletion of dsr in the chromosome of Ln. citreum CBM-D2567.