斑点叉尾鮰病毒囊膜蛋白基因ORF6和ORF10的原核表达及抗体制备

Prokaryotic Expression of Channel Catfish Herpesvirus ORF6 and ORF10 and Antibodies Preparation

为获得斑点叉尾鮰疱疹病毒ORF6和ORF10基因,设计特异性引物进行PCR扩增,成功构建pET-32a-ORF6和pET-32a-ORF10原核表达载体,经IPTG诱导后目的蛋白在大肠杆菌中成功表达并制备多抗,通过Western blot证明ORF10的抗体具有特异性,且ORF10为病毒的结构蛋白。

Primers were designed according to the channel catfish virus open reading frame 6 and 10,and both fragments of 417 bp and 456 bp were produced by PCR.The recombinant expressing plasmid pET-32a-ORF6 and pET-32a-ORF10 were constructed and subsequently expressed in E.coli after induced with IPTG.The purified proteins were used to immune rabbits separately and antibodies were obtained.Western blot analysis indicated that the polyclonal antibodies of ORF10 could react specifically with the purified CCV virions demonstrating that the ORF10 is a structural protein of CCV.