表达Rck蛋白的乳酸菌的构建  

Construction of lactic acid bacteria expressing Rck protein

作　　者：刘晶[1] 王成宇[1] 高兴[1] 陈宏亮[1] 姚心茹 杨桂连[1] 姜延龙[1] 王春凤[1] 

机构地区：[1]吉林农业大学动物科学技术学院吉林省动物微生态制剂工程研究中心,吉林长春130118

出　　处：《中国兽医科学》2016年第8期1003-1008,共6页Chinese Veterinary Science

基　　金：国家高技术研究发展计划(863)项目(2013AA102806;2011AA10A215);国家自然科学基金项目(31272552;31272541);吉林省科技发展计划项目(20111816);吉林省世行贷款农产品质量安全项目(2011-Y07)

摘　　要：为了构建侵入型乳酸菌,将来源于沙门菌的侵入型蛋白Rck及成熟Rck蛋白(m Rck)基因分别插入乳酸菌-大肠杆菌穿梭表达载体p SIP-409及锚定表达载体p SIP-409-pgs A’中,构建p SIP-409-Rck-FLAG和p SIP-409-pgs A’-m Rck-FLAG两种重组质粒,并通过电转化的方法将其电转到植物乳杆菌NC8中,通过Western-blot检测两种目的蛋白的表达情况。结果显示,Rck-FLAG在胞内表达,而m Rck-FLAG在细胞壁上表达。结果表明,两种目的蛋白均被成功表达,并且具有反应原性,为后期乳酸菌侵袭细胞试验奠定了基础。In order to construct invasive lactic acid bacteria, the invasive protein Rck and its mature form （mRck） derived from Salmonella were inserted into lactic acid bacteria-Eschericbia coli shuttle expression vector pSIP-409 and anchoring expression vector pSIP-409-pgsA＇ ,respectively,named pSIP-409-Rck-FLAG and pSIP-409-pgsA＇-mRck-FLAG. The plasmids were transformed into Lactobacillus plantarum NC8 by electroporation and the expression of proteins were detected by Western-blot. The results showed that Rck-FLAG was expressed in intracellular and mRck-FLAG in cell walls, indicating the target proteins with reactionogenicity were all successfully expressed which might lay the foundation for later invasive experiments.

关 键 词：P SIP-409 Rck 侵入型乳酸菌 质粒 

分 类 号：S852.611[农业科学—基础兽医学]