食品科学2017，Vol.38Issue(12)：15-20,6.DOI:10.7506/spkx1002-6630-201712003
同源重组法构建副干酪乳杆菌组氨酸蛋白激酶基因缺失突变株
Construction of prcK Gene Deletion Mutant of Lactobacillus paracasei by Homologous Recombination
摘要
Abstract
This study aimed to construct a histidine protein kinase gene (prcK) deletion mutant of Lactobacillus paracasei HD1.7 for providing an experiment tool for research on the function of the prcK gene.In this research,homologous recombination method was used.Plasmid pKLKRT (including prcK∷Tetr) was constructed and used to transform L.paracasei HD1.7 by eletroporation.The prcK∷Tetr in place ofprcK was integrated into the chromosome of L.paracasei HD1.7 by homologous recombination.One strain that grew only on plates with tetracycline but not on plates with ampicillin was selected.The PCR amplification and endonuclease digestion analysis indicated that plasmid pKLKRT was successfully constructed and introduced into L.paracasei HD1.7.The prcK gene deletion mutant was confirmed by PCR amplification.In conclusion,a prcK gene deletion mutant of L.paracasei HD1.7 has been successfully constructed by homologous recombination,which will lay the basis for understanding the molecular mechanism of the quorum sensing-related genes in L.paracasei HD 1.7.关键词
副干酪乳杆菌/细菌素/组氨酸蛋白激酶基因/同源重组Key words
Lactobacillus paracasei/bacteriocin/histidine protein kinase/homologous recombination分类
生物科学引用本文复制引用
岳元春,王洋,由田,高冬妮,平文祥,葛菁萍..同源重组法构建副干酪乳杆菌组氨酸蛋白激酶基因缺失突变株[J].食品科学,2017,38(12):15-20,6.基金项目
国家自然科学基金面上项目(31470537 （）
31570492) （）
黑龙江省高等学校科技创新团队(农业微生物发酵技术)项目(2012td009) （农业微生物发酵技术）
