鸭肠炎病毒SORF3基因的原核表达及其多克隆抗体的制备  

Prokaryotic Expression and Polyclonal Antibody Preparation of SORF3 Gene of Duck Enteritis Virus

作　　者：母晓宇[1] 董井泉 孟祥莉[1] 张雪莲[1] 马波[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国家禽》2012年第23期14-17,共4页China Poultry

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

摘　　要：以鸭肠炎病毒(DEV)C-KCE株基因组DNA为模板,PCR扩增获得SORF3(S3)基因,在pET-32a(+)/Rosetta(DE3)pLys系统中原核表达。S3重组蛋白经IPTG诱导,SDS-PAGE分析显示外源基因以包涵体形式表达为主,分子质量约为52ku。重组蛋白经Ni-NTA纯化后免疫新西兰白兔制备多克隆抗体,琼脂扩散法测得抗体效价为1:4。经Western blotting分析制备的多克隆抗体具有较高的特异性,间接免疫荧光试验证实制备的多克隆抗体与DEV的SORF3编码产物反应性良好,为进一步研究DEVSORF3结构与功能特性奠定了基础。With the D NA of duck enteritis virus （DEV）C-KCE as temple,SORF3 gene region was amplified by polymerase chain reaction （PCR）. Then S3 gene was expressed in pET-32a （＋）/Rosetta （DE3）pLys system. The recombinant protein of S3 induction with IPTG by SDS-PAGE analysis showed that the exogenous gene was mainly expressed as inclusion bodies,the product of S3 was about 52 ku. Rabbits were immunized with recombinant protein which were purified by Ni-NTA Purification System,and then polyclonal antibody against pET-32a-S3 was prepared. The titer of antibody tested by agar diffusion reaction was 1：4. Moreover,Western blotting revealed that the antiserum was high specificity, indirect immunofluorescence （IIF）test proved polyclonal antibody could react with DEV SORF3-encoded product,which lay foundation for further research on molecular structure and function of DEV SORF3.

关 键 词：鸭肠炎病毒 SORF3基因 原核表达 

分 类 号：S512.1[农业科学—作物学]