猪圆环病毒2型CC株ORF3基因的克隆与原核表达  

Cloning and expression of ORF3 gene of porcine circovirus 2 CC strain in E.coli

作　　者：徐廷川[1] 张亮权[1] 张得玉[1] 齐海涛[1] 张民泽[1] 张桂红[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国预防兽医学报》2012年第3期235-237,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家生猪现代农业产业体系项目(nycytx009)

摘　　要：为原核表达猪圆环病毒2型(PCV2)的ORF3编码蛋白,本研究采用PCR方法以PCV2的CC株的基因组DNA为模板扩增ORF3基因,将其克隆到原核表达载体pET-32a(+)中,转化大肠杆菌BL21(DE3),经IPTG诱导表达。SDS-PAGE结果显示,表达的重组蛋白约为31 ku,并且以包涵体形式存在;western blot分析表明,ORF3重组蛋白能够与鼠抗His标签单克隆抗体发生特异性反应,表明原核表达的ORF3重组蛋白具有良好的反应原性。To express ORF3 protein of porcine circovirus 2 （PCV2）, the ORF3 gene of PCV2 CC strain was amplified by PCR and cloned into pET32a（＋） vector to construct the recombinant plasmid of pET32a-ORF3 which was transformed into E. coli BL21 （DE3） competent cell and induced to express by 1.0 mmol/L IPTG induction, and the expressed ORF3 protein was assayed by SDS-PAGE and western blot. SDS-PAGE analysis showed that the recombinant ORF3 protein was about 31 ku, mainly in form of inclusion bodies, and western blot showed that the recombinant protein was specifically reacted with His monoclonal antibody the positive serum, indicating that the expressed protein possessed strong reactinogenicity.

关 键 词：猪圆环病毒2型 ORF3 原核表达 

分 类 号：S852.65[农业科学—基础兽医学]