致羔羊脑炎粪肠球菌XJ05 LuxS基因的克隆与原核表达  

Cloning and Prokaryotic Expression of LuxS Gene from Enterococcus faecalis XJ05 Inducing Lamb Encephalitis

作　　者：郭秉娇[1] 卢帅[1] 王熙楚[1] 郑婷婷[1] 周霞[1] 剡根强[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832000

出　　处：《动物医学进展》2014年第12期32-36,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31160507);人力资源与社会保障部出国留学人员科研启动项目(2010LX007);大学生创新创业训练项目(201210759021)

摘　　要：采用PCR技术从致羔羊脑炎粪肠球菌XJ05基因组中扩增得到LuxS基因的全序列,对其进行分析,同时与其他细菌相应基因序列及编码的氨基酸进行同源性分析;构建该基因的原核表达载体,对表达蛋白进行鉴定。结果显示,该粪肠球菌XJ05的LuxS基因全长459bp,编码152个氨基酸,具有功能序列HXXEH,核苷酸同源性与GenBank上的枯草芽胞杆菌和粪肠球菌V583同源性最高,分别为61.2%和100%;构建的重组质粒经诱导后表达分子质量为20ku的蛋白质,Western blot检测显示特异性条带。说明XJ05的luxS基因与肠球菌V583和枯草芽胞杆菌亲缘关系较近,表达的重组蛋白具有良好的反应原性。LuxS gene was amplified from the chromosomal DNA of Enterococcus faecalis XJ05 inducing lamb encephalitis with PCR, the fragments were sequenced and analyzed and compared with the corre- sponding sequences in GenBank. The prokaryotic expression vector was constructed and the recombinant proteins were analyzed by SDS-PAGE and Western blot. The full-length gene was 459 bp, the higher ho- mologies were 61.2% and 100% compared with Bocillus cereus and Enterococcus faecalis V583. The gene encoded 152 amino acids and had a high conservation HXXEH and the expressed product was 20 ku,West- ern blot showed a specific band. The results indicated that LuxS gene of XJ05 has similar relationship with that of Bocillus cereus and Enterococcus faecalis V583,and the recombinant protein had good immunoge- nicity.

关 键 词：粪肠球菌 LUXS基因 序列分析 原核表达 

分 类 号：S852.611[农业科学—基础兽医学]