Shewanella baltica luxR基因缺失菌株的构建及其功能初步研究  

Construction and analysis of luxR gene defective strain of Shewanella baltica

作　　者：揭金鑫 韩云艳 张彩丽[1] 刘尊英[1] 曾名湧[1] JIE Jin-xin;HAN Yun-yan;ZHANG Cai-li;LIU Zun-ying;ZENG Ming-yong(College of Food Science and Engineering, Ocean University of China, Qingdao 26600)

机构地区：[1]中国海洋大学食品科学与工程学院,青岛266003

出　　处：《食品科技》2018年第6期28-32,共5页Food Science and Technology

基　　金：国家自然科学基金项目(31671919)

摘　　要：细菌的luxR基因作为群体感应信号分子受体蛋白的编码基因,在细菌的群体感应及其调控的功能中发挥着重要作用。为探究冷藏凡纳滨对虾特定腐败菌波罗的海希瓦氏菌(Shewanella baltica)(WT SA03)的群体感应现象,采用PCR融合和同源重组法构建了S.baltica的luxR基因缺失菌株(ΔluxR WT SA03),对比研究了luxR基因缺失后WT SA03的生长、产硫能力及蛋白表达的差异。结果表明,经PCR扩增融合后的重组质粒在大肠杆菌中成功表达,并通过接合反应将质粒从大肠杆菌转移到野生型S.baltica WT SA03中,构建了S.baltica的luxR基因缺失株。进一步研究表明,S.baltica的luxR基因缺失对其生长并无影响,但luxR基因对其产硫能力影响显著。表明luxR基因与WT SA03的致腐能力密切相关。研究结果为深入解析S.baltica的luxR基因功能及其调控的细菌群体感应与致腐机制提供了基础。The luxR gene of bacteria encoded the receptor protein of quorum sensing molecular plays an important role in the regulation the quorum sensing behavior of bacteria. The luxR gene defective strainof Shewanella baltica（WT SA03） was constructed using PCR fusion and homologous recombination method, and the growth, H2S-producing and the protein expression of WT SA03 and ΔluxR WT SA03 were also determined. The results indicated the fragments fused by PCR were cloned into PHG1.0 plasmid, and the recombinant plasmid was successfully transducted from the E. Coli cell to the S. baltica wild-type strain by homologous recombination. The growth of ΔluxR WT SA03 has no change compared with that of WT SA03. However, H2S-producing of ΔluxR WT SA03 changed significantly, which indicated luxR gene was related to the spoilage ability of S. baltica. This research provided a basis for study of function and quorum sensing behavior of S. baltica.

关 键 词：波罗的海希瓦氏菌 PCR融合 同源重组 缺陷菌株 

分 类 号：TS254.4[轻工技术与工程—水产品加工及贮藏工程]