布鲁氏菌外膜蛋白OMP15.6表达及免疫反应原性测定

Expression and Antigenicity Identification of Brucella predicted Outer Membrane Protein 15.6

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.