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| 牛疱疹病毒I型gB基因原核表达载体的构建和高效表达 | |
| 引用本文： | 吴春涛,王建华,侯相山,霍雷,侯艳梅,牛钟相.牛疱疹病毒I型gB基因原核表达载体的构建和高效表达[J].西北农林科技大学学报(社会科学版),2007,35(1). |
| 作者姓名： | 吴春涛 王建华 侯相山 霍雷 侯艳梅 牛钟相 |
| 作者单位： | 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 |
| 基金项目： | 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。 |
| 关 键 词： | 牛疱疹病毒Ⅰ型 gB基因 原核表达 |
Construction and effective expression of the prokaryotic expression vector containing BHV-1 gB gene |
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| WU Chun-tao,WANG Jian-hua,HOU Xiang-shan,HUO Lei,HOU Yan-mei,NIU Zhong-xiang.Construction and effective expression of the prokaryotic expression vector containing BHV-1 gB gene[J].Journal of Northwest Sci-Tech Univ of Agr and,2007,35(1). | |
| Authors: | WU Chun-tao WANG Jian-hua HOU Xiang-shan HUO Lei HOU Yan-mei NIU Zhong-xiang |
| Affiliation: | WU Chun-tao~1,WANG Jian-hua~2,HOU Xiang-shan~3,HUO Lei~2,HOU Yan-mei~2,NIU Zhong-xiang~1 |
| Abstract: | The gB gene fragment of bovine herpesvirus-1 Bartha Nu/67 strain was amplified by PCR and inserted into prokaryotic expressing vector pBAD/TOPO.The recombined plasmids were then transformed 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. |
| Keywords: | bovine herpesvirus-1(BHV-1) gB gene prokaryotic expression |
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