旋毛虫T668基因真核表达载体的构建及其在BHK细胞中的表达

Construction and expression in BHK cells of eukaryotic expression plasmid with T668 gene of Trichinella spiralis

利用RT-PCR技术从中国河南猪旋毛虫(国际标准虫种编号为ISS534)新生幼虫得到T668基因,并克隆入pEGFP-N1真核表达载体中构建重组质粒,该重组质粒在脂质体介导下转染BHK细胞。EGFP标签证明质粒DNA成功转染到细胞中并得以表达,Western blotting鉴定,细胞裂解液样品中有1条约77 000的条带,可被绿色荧光抗体及猪感染旋毛虫阳性血清所识别,与预计大小一致,表明成功构建T668-pEGFP-N1真核表达质粒,并在BHK细胞融合表达,表达产物具有抗原性。

The T668 gene was amplified by RT-PCR from Trichinella spiralis NBL originated from pigs in Henan Province ISS534 and cloned into the eukaryotic expression plasmid pEGFP-N1.Then,the pEGFP-N1-T668 plasmid was transfected into BHK cells mediated by GenEscortTMⅡ.EGFP showed that the recombinant plasmid was expressed in BHK cells.Western-blotting results illustrated that there was a bands approximately in size in BHK221 cells lysate,it was recognized by the the anti-EGFP antibodies and the pig serum against T.spiralis,indicating that the T668 protein fusion expressed transiently in the cells and had antigenicity.