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.
出处
《动物医学进展》
CSCD
北大核心
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)
关键词
粪肠球菌
LUXS基因
序列分析
原核表达
Enterococcus faecalis
LuxS gene
sequence analysis
prokaryotic expression
