伪狂犬病毒gB基因的表达及单克隆抗体的制备  

Expression of the gB gene of pseudorabies virus and the preparation of its monoclonal antibody

作　　者：邱佳[1] 符芳[2] 柴政[2] 姜福成[2] 田华彬[2] 郎月坤 郑楠[2] 李曦[2] 张彦龙[1] 

机构地区：[1]黑龙江大学生命科学学院,哈尔滨150080 [2]中国农业科学院哈尔滨兽医研究所猪病分子诊断实验室,哈尔滨150001

出　　处：《黑龙江畜牧兽医》2014年第3期17-20,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：公益性行业(农业)科研专项(201203056)

摘　　要：为了研究伪狂犬病毒gB基因的原核表达并制备其单克隆抗体,试验采用含有伪狂犬病毒(PRV)gB蛋白抗原表位的基因作为模板进行扩增,得到大小为588 bp的扩增产物,将其克隆到表达载体pET-32a(+)中,转化表达菌Transetta(DE3),经诱导表达得到目的蛋白;以纯化后的重组gB蛋白为抗原,免疫Balb/c小鼠,利用淋巴细胞杂交瘤技术制备杂交瘤细胞。结果表明:表达的重组gB蛋白约为44 ku,以包涵体形式存在,具有良好的反应原性;获得的2株杂交瘤细胞均能够稳定分泌抗gB蛋白的特异性单克隆抗体(McAb)。To study the prokaryotic expression of the gB gene of pseudorabiesvirus （PRV） and prepare its monoclonal antibody, the gene containing antigenic epitope of PRV -gB protein was used as a template to amplify by PCR. A 588 bp amplified product was obtained, and then cloned into an expression vector pET -32a（ ＋ ） vector and transformed into the expression bacteria Transetta （DE3）, and the target protein was obtained by the induction expression; the purified recombinant gB protein was used as an antigen to immunize Balb/c mice, and the lymphocyte hybridoma technique was used to prepare hybridoma cells. The result showed that the recombinant gB protein was about 44 ku, which was mainly in the form of inclusion bodies, and had the good reactionogenicity; two strains of hybridoma cells obtained could stably secrete specific monoclonal antibodies （MAbs） against the gB protein.

关 键 词：伪狂犬病毒 gB蛋白 原核表达 单克隆抗体 

分 类 号：S852.659.1[农业科学—基础兽医学]