| Expression and Antigenic Characterization of the Epitope-G1 of the Bovine Ephemeral Fever Virus Glycoprotein in Pichia pastoris |
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| 作者姓名： | Fu-ying ZHENG Guo-zhen LIN Chang-qing QIU Kui-zhang YUAN Jun-ying SONG |
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| 作者单位： | 1.Key Laboratory of Animal Virology of Ministry of Agriculture，State Key Laboratory of Veterinary Etiological Biology，Lanzhou Veterinary Research Institute，Chinese Academy of Agricultural Sciences，Lanzhou 730046，China;2.Harbin Veterinary Research Institute，Chinese Academy of Agricultural Sciences，Harbin 150001，China |
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| 基金项目： | National Dairy Foundation of China (2002BA518A04) |
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| 摘 要： | The epitope-G1 gene of Bovine ephemeral fever virus(BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1.Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS115.The recombinant P.pastoris strains were selected by a G418 transformation screen and confirmed by PCR.After being induced with methanol,an expressed protein with 26 kDa molecular weight was obtained,which was much bigger than the predicted size(15.54 kDa).Deglycosylation analysis indicated the recombinant G1 was glycosylated.Western blot and ELISA tests,as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity.The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.
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| 关 键 词： | 基因表达 转基因 抗原-G1 牛流行热病毒 甲醇酵母 |
| 文章编号： | 1674-0769（2007）05-0347-06 |
| 修稿时间： | 2006-11-29 |
Expression and Antigenic Characterization of the Epitope-G1 of the Bovine Ephemeral Fever Virus Glycoprotein in Pichia pastoris |
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| Fu-ying ZHENG Guo-zhen LIN Chang-qing QIU Kui-zhang YUAN Jun-ying SONG.Expression and Antigenic Characterization of the Epitope-G1 of the Bovine Ephemeral Fever Virus Glycoprotein in Pichia pastoris[J].中国病毒学(英文版),2007,22(5):347-352. |
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| Authors: | Fu-ying ZHENG Guo-zhen LIN Chang-qing QIU Kui-zhang YUAN and Jun-ying SONG |
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| Institution: | Fu-ying ZHENG1,Guo-zhen LIN1,Chang-qing QIU1,Kui-zhang YUAN2 and Jun-ying SONG2 |
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| Abstract: | The epitope-G1 gene of Bovine ephemeral fever virus(BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1.Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS115.The recombinant P.pastoris strains were selected by a G418 transformation screen and confirmed by PCR.After being induced with methanol,an expressed protein with 26 kDa molecular weight was obtained,which was much bigger than the predicted size(15.54 kDa).Deglycosylation analysis indicated the recombinant G1 was glycosylated.Western blot and ELISA tests,as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity.The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis. |
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| Keywords: | Bovine ephemeral fever virus(BEFV) Epitope-G1 Expression Pichia pastoris |
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