捻转血矛线虫Hc38基因部分功能域的原核表达与鉴定

Expression and analysis of partial functional domains of Hc38 gene from Haemonchus contortus

为原核表达捻转血矛线虫Hc38基因的保守结构域,本研究参照GenBank中Hc38基因序列设计引物,扩增出Hc38基因的保守结构域序列,命名为HcP,将其克隆到表达载体pET32a中,并转化到大肠杆菌BL21中,用IPTG进行诱导表达。SDS-PAGE和western blot结果表明,HcP基因在大肠杆菌中高效表达,表达的重组蛋白分子量约为49.4 ku,并且表达产物能够与感染捻转血矛线虫的山羊血清产生特异的免疫印记条带,具有良好的反应原性。

To express the conserved domain sequence of Haemonchus contortus Hc38 gene（HcP）,the HcP sequence was amplified from H.contortus adult total RNA by RT-PCR with a pair of primers designed according to the cDNA available in GenBank.The HcP sequence was sub-cloned into pET32a and transformed into E.coli BL21 for expression in E.coli induced by IPTG.SDS-PAGE and western blot results showed molecular weight of the recombinant protein was about 50 ku,and recognized by positive sera against H.contortus,indicating that the recombinant protein possessed antigenic activity.