【作者】 付瑞； 贺争鸣； 邢瑞昌；

【Author】 FU Rui, HE Zheng-ming, XING Rui-cha (National Institute for the Control of Pharmaceutical and Biological Products, National Monitoring Center of Laboratory Animals (Genetics and Microbiology), Beijing 100050 )

【机构】 中国药品生物制品检定所国家实验动物微生物、遗传检测中心；

【摘要】 目的表达、纯化RHDV VP60蛋白并检测其活性,建立RHDV-VP60-ELISA检测方法。方法将重组表达质粒pET-VP60 转化入大肠杆菌BL21(DE3)菌株中,并在其中进行表达。采用脱盐以及金属鳌合亲和层析的方法对表达产物进行纯化。并用 Western blotting方法检测表达产物的抗原性。结果在大肠杆菌中以包涵体形式高效表达了重组RHDV VP60蛋白,经过纯化使蛋白纯度达到98％,经Western blotting方法检测表明该重组蛋白有一定的抗原性。结论通过原核表达可以得到具有抗原性的重组RHDV VP60蛋白。更多还原

【Abstract】 Objective To express and purify the RHDV VP60 protein, then detect its activity, and establish the RHDV-VP60-ELISA method. Methods The vector pET-VP60 was transformed into E. coli BL21(DE3). The recombinant protein was expressed. Then, the protein was purified by desalting and affinity chromatography, and its activity was analysed by Western blotting. Results The recombinant RHDV VP60 protein has been expressed in E.coli, and its purity can reach 98%. The activity of the recombinant protein was proved through Western blotting. Conclusion The recombinant RHDV VP60 protein can be expressed in prokaryotic system. And the protein has antigenicity.更多还原