植物乳杆菌luxS基因敲除载体的构建与应用

o construct the Lactobacillus plantarum KLDS 1.0391 homologous recombination vector and achieve gene knockout. Using genome as template, the flanking sequences of luxS gene were amplified by PCR and cloned into pMD 19-T Simple Vector. After colony PCR identification, these two recombination vectors pMD19-T Simple-luxSL、pMD19-T Simple-luxSR were digested by different restriction enzymes and cloned into pNZ5319. After identification, the homologous recombination vector pNZ5319-luxS was transformed into E.coli DH5α.The L.plantarum KLDS1.0391 luxS knockout strain was constructed by using the method of electroporation. The results showed that the luxS gene knockout vector pNZ5319-luxS was constructed successfully, and the sequencing results showed that one of the two luxS genes was successfully knocked out. The important role of luxS in metabolism and quorum sensing of L. plantarum KLDS1.0391 will be based on this work.