| 摘要: |
| 用PCR法扩增了牛疱疹病毒Ⅰ型Bartha Nu/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TOP10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-Arabinose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-Arabinose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 |
| DOI： |
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| 基金项目:天津市应用基础研究计划项目(YFJMJC12000) |
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| Construction and effective expression of the prokaryotic expression vector containing BHV-1 gB gene |
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| 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. |
| Key words: bovine herpesvirus-1 (BHV-1) gB gene prokaryotic expression |
