| 犬瘟热病毒水貂株H与F蛋白基因原核表达重组质粒的构建及表达 |
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| 引用本文： | 苏凤艳,温铁峰,宗颖,王全凯.犬瘟热病毒水貂株H与F蛋白基因原核表达重组质粒的构建及表达[J].华南农业大学学报,2012,33(3):417-420. |
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| 作者姓名： | 苏凤艳 温铁峰 宗颖 王全凯 |
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| 作者单位： | 吉林农业大学中药材学院,吉林长春,130118 |
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| 基金项目： | 吉林省自然科学基金，吉林省教育厅项目 |
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| 摘 要： | 将克隆质粒pMD-18T-H和pMD-18T-F分别进行双酶切,获得纯化的H基因和F基因,在T4DNA连接酶的作用下,亚克隆到原核表达载体pET28a(+)上,获得原核重组质粒pET28a-H和pET28a-F.将原核重组质粒转化大肠埃希菌Escherichia coli Rosetta2(DE3)中进行表达,SDS-PAGE结果显示H基因和F基因分别表达相对分子质量约为31 400和38 200的融合蛋白,Western-blot检测结果显示,表达蛋白均可与CDV标准阳性血清呈阳性反应,表明原核表达的CDV H蛋白和F蛋白在反应原性上具有与天然H蛋白和F蛋白同样的特性,可作为CDV诊断用抗原,为CDV的免疫预防研究奠定基础.
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| 关 键 词： | 犬瘟热病毒 H基因 F基因 原核表达 |
| 收稿时间： | 2011/12/27 0:00:00 |
Construction and Expression of Protokaryon Expression Plasmid for H and F Genes of Canine Distemper Virus Isolated from Mink |
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| SU Feng-yan,WEN Tie-feng,ZONG Ying and WANG Quan-kai.Construction and Expression of Protokaryon Expression Plasmid for H and F Genes of Canine Distemper Virus Isolated from Mink[J].Journal of South China Agricultural University,2012,33(3):417-420. |
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| Authors: | SU Feng-yan WEN Tie-feng ZONG Ying and WANG Quan-kai |
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| Institution: | (College of Chinese Medicinal Materials,Jilin Agricultural University,Changchun 130118,China) |
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| Abstract: | H and F genes were obtained by digesting the cloning plasmid pMD-18T-H and pMD-18T-F respectively.The recombinant protokaryon expression plasmids pET28a-H and pET28a-F were constructed by sub-cloning the objective gene into pET28a(+) vector.Recombinant protokaryon expression plasmids pET28a-H and pET28a-F were transformed into Escherichia coli Rosetta2(DE3) cell.Target genes were successfully expressed by SDS-PAGE detection.The size of the recombinant fusion proteins was 31 400(H) and 38 200(F) respectively.The recombinant fusion proteins showed positive reaction to anti-CDV canine serum by Western-blot detection. |
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| Keywords: | canine distemper virus H gene F gene protokaryon expression |
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