猪圆环病毒2型ORF2基因及片段的原核表达  

Prokaryotic Expression of ORF2 Gene and Its Four Different Fragments of Porcine Circovirus Type 2

作　　者：屈素洁[1] 黄伟坚[1] 冯建远[1] 朱芸[1] 陈琼[1] 谢丽华[1] 

机构地区：[1]广西大学动物科学技术学院,广西南宁530005

出　　处：《实验动物科学》2007年第6期89-92,共4页Laboratory Animal Science

基　　金：国家科技攻关计划(2004BA717B-01-03);广西科学研究与技术开发项目(桂科攻0537008-3);广西壮族自治区科技攻关项目(No:0537022-1桂科能No:0630006-6B)

摘　　要：目的表达猪圆环病毒2型ORF2抗原,为研制猪圆环病毒2型血清学诊断方法以及生物制品生物安全性检验,提供技术支持。方法根据猪圆环病毒2型(PCV2)LC株序列,设计合成5对引物,采用PCR方法从全序列重组质粒PCV2-LC中扩增出了ORF2基因和4个不同ORF2基因片段,分别将其克隆到原核表达载体PET-32a上,再分别将各个重组表达质粒转化到大肠杆菌BL21中,用终浓度1 mmol/LIPTG诱导。结果表达产物以包涵体的形式存在,经Western-blot检测表明,表达的重组蛋白ORF2b、ORF2c、ORF2d能够被PCV2阳性血清所识别,具有良好的抗原性。表达的重组蛋白为ELISA诊断试剂盒的研制奠定了基础。Based on the porcine circovirus type 2 （PCV2）LC strain,5 pair of primers were designed according to the Luchuan sequence, the ORF2, ORF2a, ORF2b, ORF2c and ORF2d gene were amplified from the PCV2-LC plasmids by PCR, and inserted into the prokaryotic expression vector PET-32a. The constructed recombinant expression plasmids were transformed into the receptive cells of E. coli BL21 and induced by IPTG with a finial concentration 1 mmol/L, then the recombinant protein was expressed in the form of inclusion bodies. The protein expression could react with PCV2 positive serum by Western-blot, sharing a good antigenicity. This study will provide a good fundament for development of PCV2 diagnostic ELISA diagnosis assay kit.

关 键 词：猪圆环病毒2型 ORF2基因 原核表达 

分 类 号：S851.347[农业科学—预防兽医学]