鸭肠炎病毒US10基因的原核表达及抗血清的制备  

Prokaryotic Expression and the Antiserum Preparation of Duck Enteritis Virus US10 Gene

作　　者：张蕾[1] 董井泉 母晓宇[1] 马波[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学院,哈尔滨150030

出　　处：《吉林农业大学学报》2013年第1期58-62,71,共6页Journal of Jilin Agricultural University

基　　金：高等学校博士学科点专项科研基金资助课题(20102325110004);东北农业大学博士启动基金项目(2009)

摘　　要：以鸭肠炎病毒(DEV)C-KCE株基因组DNA为模板,PCR扩增获得US10基因。构建了其原核表达载体pET-32a-US10,经1.0 mmol/L IPTG诱导,目的蛋白在Rosetta(DE3)pLys宿主菌中获得了表达。SDS-PAGE电泳结果显示目的蛋白以包涵体形式表达,与预期大小相符。利用亲和层析法纯化目的蛋白,以纯化的蛋白为免疫原制备其抗血清,琼脂扩散法测得抗体效价为1∶4,间接ELISA法测定抗体效价为1∶102 400。间接免疫荧光试验证实制备的抗血清可特异性识别鸭肠炎病毒的US10基因编码产物。The US10 gene was expressed in pET-32a（＋）/Rosetta（DE3） pLys system using the DNA of Duck enteritis virus（DEV） C-KCE as temple,US10 gene ORF was amplified by polymerase chain reaction（PCR）.Then the recombinant protein of US10 gene induction with IPTG by SDS-PAGE analysis showed that the object protein is mainly expressed as inclusion bodies.Using the Ni-NTA Purification System,the recombinant protein immunized to rabbits was purified to make the antiserum.The titer of the antiserum tested by agar diffusion reaction was 1∶4.The titer of the antiserum tested by ELISA was 1∶102 400.The prepared antiserum has a high specificity with the western-blot analysis,and the indirect immunofluorescence（IIF） test proved that the antiserum could react with DEV US10 encoded product,which lays a foundation for further research on molecular structure and function.

关 键 词：鸭肠炎病毒 US10基因 表达 

分 类 号：S852.65[农业科学—基础兽医学]