| I型鸭肝炎病毒VP1、3D基因克隆及其在大肠杆菌中的表达 |
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| 引用本文： | 孔留五,罗玉均,张桂红,陈建红,徐小芹,廖 明,康艳梅,何逸民.I型鸭肝炎病毒VP1、3D基因克隆及其在大肠杆菌中的表达[J].微生物学通报,2008,35(7):1068-1071. |
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| 作者姓名： | 孔留五 罗玉均 张桂红 陈建红 徐小芹 廖 明 康艳梅 何逸民 |
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| 作者单位： | 华南农业大学兽医学院 广州 510642;(1. 华南农业大学兽医学院 广州 510642)(2. 佛山科学技术学院生命科学学院 南海 528231);华南农业大学兽医学院 广州 510642;佛山科学技术学院生命科学学院 南海 528231;华南农业大学兽医学院 广州 510642;华南农业大学兽医学院 广州 510642;华南农业大学兽医学院 广州 510642;华南农业大学兽医学院 广州 510642 |
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| 基金项目： | 广东省自然科学基金(No. 5006678) |
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| 摘 要： | 根据GenBank中的I型鸭肝炎病毒全基因序列设计了扩增I型鸭肝炎病毒VP1、3D基因的引物, 用该特异性表达引物从I型鸭肝炎病毒cDNA模板中扩增得到目的基因VP1、3D, 用相同的限制性内切酶酶切目的基因和表达载体pET32a后构建重组表达载体, 转化宿主BL21(DE3), 用不同浓度的IPTG诱导VP1、3D基因的表达, 收集菌液进行SDS-PAGE电泳, Western-blotting分析蛋白免疫原性。结果表明, VP1、3D在大肠杆菌中表达量较高, 表达产物的分子量约为48 kD、68 kD, 并能被兔抗DHV-1血清所识别。I型鸭肝炎病毒VP1、3D蛋白在大肠杆菌中表达产物具有免疫原性。
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| 关 键 词： | I型鸭肝炎病毒 VP1基因 3D基因 克隆表达 |
Cloning of VP1 and 3D Gene of Duck Hepatitis Virus 1 (DHV1) and Its Expression in Escherichia coli |
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| KONG Liu-Wu,LUO Yu-Jun,ZHANG Gui-Hong,CHEN Jian-Hong,XU Xiao-Qin,LIAO Ming,KANG Yan-Mei and HE Yi-Min.Cloning of VP1 and 3D Gene of Duck Hepatitis Virus 1 (DHV1) and Its Expression in Escherichia coli[J].Microbiology,2008,35(7):1068-1071. |
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| Authors: | KONG Liu-Wu LUO Yu-Jun ZHANG Gui-Hong CHEN Jian-Hong XU Xiao-Qin LIAO Ming KANG Yan-Mei and HE Yi-Min |
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| Institution: | College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642;(1. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642) (2. College of Life and Science, Foshan University, Nanhai 528231);College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642;College of Life and Science, Foshan University, Nanhai 528231;College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642;College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642;College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642;College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642 |
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| Abstract: | In this study, two special primer pair containing EcoR V and Xho I according to complete genome of duck hepatitis virus 1 (DHV1) were designed to amplify VP1 and 3D genes from cDNA of DHV1. The target genes VP1 and 3D were subcloned into PET32a vector digested by EcoR V and Xho I respectively. Then the recombinant plasmids were transfected into Escherichia coli BL21(DE3) for VP1and 3D expression. The bacteria containing PET32a-VP1 and PET32a-3D were collected and examined by SDS-PAGE and western-blotting. Result showed that the VP1 and 3Dprotein were expressed in E. coli and the amount of expression was higher. Molecular weight of the protein was 48 kD, 68 kD. The protein can be recognized by DHV1antibody. This study showed that the protein VP1 and 3D have antigenicity. |
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| Keywords: | Duck hepatitis virus 1 VP1gene 3Dgene Cloning and expression |
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