棉铃虫病毒gp41基因的克隆表达及抗体制备

根据棉铃虫单核衣壳核多角体病毒(Helicoverpa armigerasingle nucleocapsid nucleopolyhedrovirus,HaSNPV)gp41基因的序列,设计引物,引入适当的酶切位点,通过PCR的方法扩增目的片段。将扩增出的基因片段克隆至原核表达载体pET-28a,构建重组质粒并转化至大肠杆菌中,经IPTG诱导表达。纯化蛋白产物并免疫家兔产生抗血清。该抗血清可与原核表达的His-GP41融合蛋白及在感染的昆虫细胞中表达的GP41蛋白发生特异性免疫反应。该抗体的获得为深入研究GP41的功能提供了基础。

Molecular Cloning, Expression and Antibody Preparation of the gp41 Gene of HaSNPV

The PCR product of Helicoverpa armigera single nucleocapsid nucleoployhedrvirus(HaSNPV)gp41 gene was cloned to a prokaryotic expression vector pET-28a.After inducing with IPTG,the gp41 gene was successfully expressed in E.coli DH5α.The protein was purified and injected into rabbit to raise polyclonal antiserum.Western blot analysis using this antiserum showed that the specific reaction to GP41 protein expressed in both prokaryotic cells with His fusion and viral infected insect cells.The antibody provides a basement for further studying on the function of GP41 protein.