大肠杆菌“菌影”的制备  

Generation of Escherichia coli ghosts

作　　者：彭玮[1,2] 王聃[2] 刘思国[2] 常月红[2] 王春来[2] 刘慧芳[2] 司薇[2] 赫明雷[2] 杜艳芬[2] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室动物细菌病研究室,哈尔滨150001

出　　处：《东北农业大学学报》2009年第6期85-88,共4页Journal of Northeast Agricultural University

基　　金：国家高技术研究发展计划(2006AA10A206)

摘　　要：通过PCR扩增噬菌体PhiX174的裂解基因E,将该基因连接到含有PR/cI启动阻遏系统的pBV220载体中,从而使裂解基因E和启动阻遏系统λPL/PR-cI857串联成为温度敏感的裂解盒,构建溶菌质粒pBV-E。采用CaCl2法将其转入大肠杆菌BL21(DE3),含有裂解质粒的大肠杆菌在28℃条件下生长到对数生长期,然后升温到42℃诱导其溶解,制成了大肠杆菌菌影。电镜观察发现菌影形态完整,内容物全部被释放到胞外。In the present study, lytic gene E from PhiX174 was amplified by PCR and inserted into pBV220 vector which included the λPI/PR-cI857 regulatory system. Lytic gene E was connected in series with the λPI/PR-cI857 regulatory system to form the temperature-sensitive lysis case that was in constructed bacteriolysis plasmid pBV-E. The plasmid pBV-E was transformed into E. coli BL21（DE3） by calcium chloride method. E. coli with the gene E was grown in a waterbath shaker at 28 ℃ under aerobic conditions until mid log-phase. Then the culture was transferred to 42 ℃ water-bath shaker. Finnally, E.coli ghosts were produced, which were in intact forms under transmission electron microscope, and their contents were released to extra cellular region. This experiment laid a foundation of study on bacterial ghost as a novel vaccine and form of adjuvant.

关 键 词：大肠杆菌 菌影 裂解基因E 

分 类 号：S852.612[农业科学—基础兽医学] Q785[农业科学—兽医学]