马立克病毒UL36^(USP)蛋白的表达及其抗体制备  

Expression and polyclonal antibody preparation of UL36^(USP) protein encoded by MDV

作　　者：郑海南[1] 邓晨[1] 王梦云[1] 金雯[1] 吴山力[2] 吕岩[1] 于子阳[3] 艾永兴[1] 

机构地区：[1]吉林大学动物科学学院,吉林长春130062 [2]吉林大学白求恩医学院,吉林长春130021 [3]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医学报》2014年第5期712-716,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31272528);教育部新世纪优秀人才支持计划资助项目(NCET-12-0232);教育部留学回国人员科研启动基金资助项目

摘　　要：UL36USP是马立克病毒UL36基因编码的蛋白UL36上具有去泛素化酶活性的N-端部分片段(UL36USP),本试验以提取MDCC-MSB1细胞的基因组为模板通过PCR获得目的基因,将UL36USP基因克隆入pMD18T载体,测序正确并酶切鉴定后,将目的基因亚克隆到pET28a原核表达载体,构建重组质粒pET28a-UL36USP。将重组质粒转化BL21(DE3)E.coli中,IPTG诱导表达并使用Ni-NTA Agarose亲和层析进行纯化得到目的蛋白。应用纯化后的UL36USP免疫獭兔制备多克隆抗体。使用昆虫细胞表达系统表达的UL36片段检测多克隆抗体的特异性。结果显示成功制备特异性的UL36USP抗体。UL36^USP,N-terminal fragment of UL36 protein encoded by MDV UL36 gene, has deubiquitination activity. In current study,UL36^USP gene was amplified from MDCC-MSB1 genome by PCR and cloned into pET28a expression vector. The recombinant plasmid was transformed into BL21（DE3） competent cells and induced with IPTG for expression. The protein was purified by Ni-NTA purification system. Rex rabbit were immunized with the purified UL36^USP for the generation of polyclonal antibody. The specificity of the antibody was determined by UL36^USP protein expressed by Bac-to-Bac system.

关 键 词：UL36基因 马立克病毒 去泛素化 多克隆抗体 

分 类 号：S852.65[农业科学—基础兽医学]