摘要
表达羊布鲁氏菌16M预测外膜蛋白OMP15.6,探索其作为诊断抗原的可能性。采用降落PCR方法,从羊布鲁氏菌16M基因组DNA中扩增出423bp的基因片段,将该片段克隆于原核表达载体PGEX-4T-2,构建重组表达载体。经IPTG诱导,SDS-PAGE检测,结果表明,在大肠杆菌中成功表达了外膜蛋白OMP15.6。经West-ern-blotting检测,该蛋白能与豚鼠抗布鲁氏菌阳性血清发生特异性免疫反应,为其之后的抗原性以及免疫原性研究奠定了良好的基础。
In this study, the outer membrane protein (OMP) 15.6 of Brucella mellitensis 16M was expressed, and the posiibility of using this outer membrane proteins as diagnostic antigen was explored. The gene encoding OMP15.6 was obtained by touch-down PCR from Brucella mellitensis 16M genome, and cloned into the expressing vector PGEX-4T-2. The SDS-PAGE showed that the OMP15.6 was sucssessfully expressed in E.coli. Western-blotting showed the OMP15.6 can react with hamster semrum agansit Brucella mellitensis 16M. The research provided the foundation for further study of immunogenesity and antigennisity of OMP15.6.
出处
《中国动物检疫》
CAS
2010年第9期29-30,共2页
China Animal Health Inspection
基金
现代农业(肉牛)产业技术体系建设专项资金
国家科技部科技支撑计划项目(2006BAD04A05-03)