植物乳杆菌KLDS1.0391 luxS基因敲除载体的构建与应用  

Construction and application of Lactobacillus plantarum KLDS1.0391 luxS gene konckout vector

作　　者：郑慧琦[1] 庞雪辉[1] 朱宗涛[1] 孟祥晨[1] 

机构地区：[1]东北农业大学乳品科学教育部重点实验室,哈尔滨150030

出　　处：《中国科技论文》2016年第6期694-698,共5页China Sciencepaper

基　　金：高等学校博士学科点专项科研基金资助项目(20122325110017);哈尔滨市杰出青年人才基金资助项目(2014RFYXJ006)

摘　　要：构建植物乳杆菌KLDS1.0391的luxS基因敲除载体并实现基因敲除。以植物乳杆菌KLDS1.0391的基因组为模板,采用PCR方法扩增luxS侧翼序列luxSL、luxSR并分别克隆至pMD19-T simple中,鉴定正确后酶切,与敲除载体pNZ5319连接,鉴定正确后转入大肠杆菌DH5α中保存。采用电转化方法将敲除载体导入植物乳杆菌KLDS1.0391,构建luxS基因敲除菌株。结果表明,植物乳杆菌luxS基因敲除载体pNZ5319-luxS构建成功,测序结果证实该菌中存在2个luxS基因且成功敲除1个。所得结果为进一步研究luxS基因在植物乳杆菌KLDS1.0391的代谢和群体感应中的重要作用奠定了基础。Constructing the Lactobacillus plantarum KLDS1. 0391 homologous recombination vector and achieving gene knockout were investiged. Using genome as template, the flanking sequences of luxS gene were amplified by PCR and cloned into vector pMD19-T simple. After colony PCR identification, the two recombination vectors pMD19-T simple-luxSL and pMD19-T simpleluxSR were digested by different restriction enzymes and cloned into pNZ5319. After identification, the homologous recombina-tion vector pNZ5319-lwxS was transformed into E. coli DH5a. The L. plantarum KLDS1. 0391 luxS gene knockout strain was constructed by using the method of electroporation The results show that the luxS gene knockout vector pNZ5319-luxS is con-structed successfully, and the sequencing results indicate that one of the two luxS genes is successfully knocked out. The important role of luxS in metabolism and quorum sensing of L. plantarum KLDS1. 0391 will be based on this work.

关 键 词：食品科学 植物乳杆菌 LUXS基因 载体构建 基因敲除 

分 类 号：Q78[生物学—分子生物学]