口蹄疫病毒VP2基因的真核表达及其产物抗原性的检测

Eukaryotic expression of VP2 gene of foot-and-mouth disease virus and examination of expression product antigenicity

利用PCR技术,扩增出口蹄疫病毒(FMDV)VP2基因,并克隆到杆状病毒转移载体pBlueBacHis2A上。用重组质粒pB-VP2与重组病毒同时转染sf9昆虫细胞,获得了重组病毒。经过蚀斑筛选纯化后,感染sf9细胞,表达VP2融合蛋白,分子质量为33 ku左右。以牛抗O型FMDV血清为第一抗体,通过Western-blotting和Dot-ELISA鉴定,说明VP2基因在真核表达系统中获得正确表达,且可以与牛抗O型FMDV血清发生特异性反应。

The FMDV VP2 gene was amplified by the VP2 gene primers. The amplified fragment was cloned into the vector pMD18-T. The recombinant plasmid was transformed into the E. coli TG1, and digested with Pst Ⅰ and Kpn Ⅰ , then cloned into transfer vector pBlueBacHis2A. The recombinant euka- ryotic transfer vector pBlueBacHis2A-VP2 was used to transfect into insect cell sf9. After plaque scan and two times amplification of the virus stocks, the target protein was expressed in insect cell sf9, the molecular weight was 33 ku. The Fusion protein antigenicity was authenticated through Western-blotting and Dot-ELISA by using cattle antisera as first antibody.