B组轮状病毒WH-1株nsp2基因的原核表达与抗体制备

Prokaryotic Expression of nsp2 Gene of Group B Rotavirus and Preparation of Its Antiserrum

根据B组轮状病毒(GBRV)WH-1株nsp2基因的全序列,设计引物,用PCR的方法扩增得到nsp2基因的编码区.将其片段克隆到原核表达载体pGEX-KG上,经IPTG诱导,在E.coliDH5α菌株中得到高效表达.表达的GST融合蛋白大小为61×103,经SDS-PAGE分离纯化,免疫小白鼠制备了多克隆抗体.抗体经1∶3 000倍稀释后用于Western Blot分析,获得特异性显色信号.所表达的蛋白和制备的抗体可用于蛋白结构和功能的进一步研究.

The ORF of nsp2 gene was amplified by PCR from the nsp2 gene sequence of Group B rota virus （GBRV）. The PCR product was cloned into the expression vector pGEX-KG. After IPTG induction,the E. coli DH5α strain containing the recombinant plasmid expressed fusion protein with molecular weights of 61×10^3 , which was in agreement with the prospectation. The purified recombinant protein was suitable to be used to immunize the mouse. Western Blot analysis using the muhiclonal antibody derived from the mouse indicated that these antibodies could react with the target protein and were suitable to be used for further functional analysis of the nsp2 gene.