小反刍兽疫病毒M蛋白主要抗原表位区的原核表达及鉴定

Prokaryotic expression and identification of M protein main epitopes of peste des petits ruminants virus

为了制备小反刍兽疫病毒(PPRV)特异性单抗及临床抗体水平监测,通过在线软件分析PPRVNigeria75/1株M蛋白可能包含的B细胞线性表位位点,结合可溶性预测,设计1对引物,扩增303bp目的基因。将目的基因克隆至原核表达载体pET-32a(+)中,用IPTG诱导表达,纯化,SDS-PAGE分析和Western-blot鉴定结果显示,所构建的重组蛋白以可溶性形式高效表达并具有良好的反应原性,为制备特异性单抗及包被抗原用于PPRV抗体水平监测奠定了一定基础。

To prepare peste des petits ruminants virus（PPRV）-specific monoclonal antibody and moni- tor the clinical antibody level of PPRV, putative B-cell linear epitope sites of M protein of PPRV Nigeria75/1 strain was analysed by online software. A pair of primers was designed to amplify a 303 bp tar- geting gene fragment which was then cloned into the prokaryotic expression vector pET-32a（＋）. The re- combinant protein was expressed in Escherichia coli BL21 （DE3） by IPTG induction and purified. SDS- PAGE showed that the recombinant was expressed in the solube form with high expression level and Wes- tern-blot anal protein level of could ysis indicated that the purified M protein possessed good reactinogenicity. The expressed M be used for the preparation of specific monoclonal antibodies and surveillance of antibody PPRV.