烟夜蛾滞育激素基因在粟酒裂殖酵母中的表达  

Expression of Diapause Hormone Gene from Helicoverpa assulta in Schizosaccharomyces pombe

作　　者：赵特[1] 罗梅浩[1] 郭线茹[1] 原国辉[1] 

机构地区：[1]河南农业大学植物保护学院,河南郑州450002

出　　处：《河南农业大学学报》2008年第4期423-426,共4页Journal of Henan Agricultural University

基　　金：河南省自然科学基金项目(0411033300)

摘　　要：利用设计的特异性引物通过RT-PCR的方法获得了烟夜蛾滞育激素(Diapause hormone,DH)基因,构建了真核表达载体pESP-2-DH,并在粟酒裂殖酵母SP-Q01中进行了诱导表达,对表达产物进行了SDS-PAGE电泳,获得了与谷胱甘肽S-转移酶(Glutathione S-tranferases,GST)融合表达的蛋白,分子量约48 kDa,与预测的融合蛋白分子量大小相当。对表达的融合蛋白进行Western blotting检测,结果表明,与GST抗体产生了很强的交叉反应,表明融合蛋白得到了表达.The cDNA encoding diapause hormone gene from Helicoverpa assulta was cloned by RT-PCR amplification using gene specific primers and then it was subcloned into expression vector pESP-2. The fusion protein was expressed in Schizosaccharomyces pombe cell and was identified by SDS polyacrylamide gel electrophoresis and Western blotting. The molecular mass of fusion protein with GST Tag was found to be 48 kDa, approximately equal to the forcasted molecular mass of the fusion protein. Results of Western blotting testing of the expressed fusion protein shows that it could have strong immune crossreaction with GST antigens,showing that the fusion protein was expressed.

关 键 词：烟夜蛾 滞育激素 RT-PCR 真核表达 融合蛋白 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]