猪戊型肝炎病毒ORF2基因的分段原核表达及抗原表位的初步鉴定  

Prokaryotic Expression and Preliminary Identification of Antigenic Epitope of the Different Fragments of ORF2 Gene of Swine Hepatitis E Virus

作　　者：古洪浪[1] 梁焕斌[1] 纪方晓 周沛[1] 邓胜朝 曹振鹏[1] 王衡[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：《中国兽医杂志》2015年第9期25-27,共3页Chinese Journal of Veterinary Medicine

基　　金：现代农业产业技术体系建设专项资金资助(CARS-36);高等学校博士点学科专项科研基金(20114404120010;2-0104404110009);广东省教育厅育苗工程(2012LYM_0031)

摘　　要：为了初步鉴定猪戊型肝炎病毒（sw HEV）ORF2蛋白的抗原表位,通过设计两对引物,PCR扩增ORF2-1和ORF2-2两段基因片段,使用p MD18-T载体克隆,双酶切后构建重组质粒p ET32a-0RF2-（1,2）,转化大肠杆菌BL21后进行IPTG诱导和SDS-PAGE凝胶电泳,最后用4株ORF2单抗对其进行Western Blot来初步鉴定其抗原表位。结果表明,分别获得约为36 ku和35 ku的两个目的蛋白,均以不可溶形式存在,抗原表位初步定位于ORF2-1这段蛋白中的414~523 aa。To identify the antigenic epitope of the ORF2 protein of swine hepatitis E virus,the ORF2-1 and ORF2-2 frag-ments were respectively amplified by designing two pairs of specific primers.The two fragments were cloned into p MD18-T vector.The recombinant plasmids p ET32a-ORF2-1,2 were constructed and transformed into E.coli BL21.After IPTG induction,the pro-tein was analyzed by SDS-PAGE and Western-blot. Its antigenic epitope of the ORF2 was preliminarily identified by four ORF2 monoclonal antibodies.The result showed that the two target proteins were obtained successfully and they were 36 ku and 35 ku re-spectively.Its antigenic epitope was preliminarily identified as the 414~523 aa of the ORF2-1 protein.

关 键 词：猪戊型肝炎病毒 ORF2基因 原核表达 抗原表位 

分 类 号：S852.651[农业科学—基础兽医学]