牛疱疹病毒I型gB基因原核表达载体的构建和高效表达  

Construction and effective expression of the prokaryotic expression vector containing BHV-1 gB gene

作　　者：吴春涛[1] 王建华[2] 侯相山[3] 霍雷[2] 侯艳梅[2] 牛钟相[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]天津出入境检验检疫局,天津300456 [3]东营职业学院农业工程系,山东东营257091

出　　处：《西北农林科技大学学报（自然科学版）》2007年第1期19-23,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：天津市应用基础研究计划项目(YFJMJC12000)

摘　　要：用PCR法扩增了牛疱疹病毒Ⅰ型B artha N u/67株gB基因,将其插入原核表达载体pBAD/TOPO中构建重组质粒pBAD-gB。将pBAD-gB质粒转化大肠杆菌TOP 10后进行了诱导表达,对表达产物进行了纯化和抗原性检测,并通过改变诱导剂L-A rab inose的浓度和诱导时间对诱导表达条件进行了优化。结果表明,重组质粒pBAD-gB构建成功,gB蛋白获得了高效表达,并以包涵体形式存在,纯化后gB蛋白的纯度达90%以上,gB蛋白抗原性良好,最佳诱导表达条件:L-A rab inose终浓度为0.2 g/L,诱导时间为5 h。The gB gene fragment of bovine herpesvirus-1 Bartha Nu/67 strain was amplified by PCR and inserted into prokaryotie expressing vector pBAD/TOPO. The recombined plasmids were then trans- formed into E. coli TOP10 for expression. The expression was optimized with proper inducing conditions of 0. 2 g/L L-Arabinose and 5 hours induction. The SDS-PAGE assay showed that the recombined plasmids could be expressed in inclusion body at a high level when induced with L-Arabinose. The purity of fusion proteins which was purified by magnehis protein purification system was more than 90%. Western-blot showed the good antigenicity of the target proteins.

关 键 词：牛疱疹病毒Ⅰ型 GB基因 原核表达 

分 类 号：S851.347.101[农业科学—预防兽医学]