兔出血症病毒主要结构蛋白基因的克隆、原核表达及其免疫原性鉴定

本试验采用RT-PCR方法扩增了兔出血症病毒(rabbit haemorrhagic disease virus,RHDV)衣壳蛋白VP60基因。PCR产物纯化后连接T载体,提取质粒后,进行序列测定,将测序正确的纯化产物双酶切,克隆入原核表达载体pET28b(+)中,构建原核表达载体。将鉴定正确的重组质粒转化表达宿主菌E.coli Rosetta^TM,用IPTG诱导培养重组表达菌。经SDS-PAGE分析,在分子质量62.0 ku处可见明显的表达带,经Western blotting检测,约在62.0 ku处出现特异性的反应带。结果表明,表达蛋白具有良好的抗原性。

Cloning,Expression and Immunogenicity of Major Structural Protein Gene of Rabbit Haemorrhagic Disease Virus

The VP60 gene of rabbit haemorrhagic disease virus was cloned by RT-PCR and sequenced after purification,and then the amplied fragment was linked into prokaryotic expression vector pET28b(+).The recombinant plasmid was transformed into E.coli JM109.The identification was right,the recombinant plasmid was transformed into host strain E.coli RosettaTM,and induced by IPTG.The expressed protein of 62.0 ku was identified by SDS-PAGE and Western blotting.