| 抗伪狂犬病病毒gC抗体制备及其gC蛋白表达 |
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| 引用本文： | 刘庆伟,邱亚峰,王晓杜,郭林,刘超,沈阳,李向东,单虎,马志永.抗伪狂犬病病毒gC抗体制备及其gC蛋白表达[J].中国畜牧兽医,2011,38(3):108-111. |
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| 作者姓名： | 刘庆伟 邱亚峰 王晓杜 郭林 刘超 沈阳 李向东 单虎 马志永 |
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| 作者单位： | (1.中国农业科学院上海兽医研究所兽医公共卫生实验室，上海 200241；2.青岛农业大学动物科技学院，山东青岛 266109) |
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| 基金项目： | 国家自然科学基金青年基金，中央级公益性科研院所基本科研业务费专项资金项目 |
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| 摘 要： | 用纯化的重组蛋白His-gCN813制备抗PRV-gC抗体，并分析伪狂犬病病毒（pseudorabies virus,PRV）gC蛋白在真核细胞的表达情况。本研究以提取的病毒基因组为模板，PCR克隆PRV gC全长基因，构建gC真核表达质粒p3xFLAG-gC，在真核细胞中表达gC基因；纯化蛋白His-gCN813制备的抗体,Western blotting和间接免疫荧光（IFA）检测到p3xFLAG-gC转染真核细胞和PRV感染细胞中的gC蛋白。结果表明本试验成功构建了PRV gC基因真核表达系统，获得了特异性抗PRV gC抗体，重组gC真核表达质粒p3xFLAG-gC转染Vero细胞，表达的重组蛋白gC为72 ku，PRV感染Vero细胞，表达的gC蛋白为55、72和94 ku，主要定位在细胞浆，gC蛋白可以作为PRV感染的指示分子，为下一步研究PRV和宿主相互作用及PRV的复制奠定了基础。
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| 关 键 词： | 伪狂犬病病毒 抗 PRV gC抗体 Western blotting |
Generation of Anti-PRV gC Antibodies and Analysis of PRV gC Expression |
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| LIU Qing-wei,QIU Ya-feng,WANG Xiao-du,GUO Lin,LIU Chao,SHEN Yang,LI Xiang-dong,SHAN Hu,MA Zhi-yong.Generation of Anti-PRV gC Antibodies and Analysis of PRV gC Expression[J].China Animal Husbandry & Veterinary Medicine,2011,38(3):108-111. |
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| Authors: | LIU Qing-wei QIU Ya-feng WANG Xiao-du GUO Lin LIU Chao SHEN Yang LI Xiang-dong SHAN Hu MA Zhi-yong |
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| Institution: | （1.Department of Veterinary Public Health，Shanghai Veterinary Research Institute，Chinese Academy of Agricultural Sciences，Shanghai 200241，China; 2.College of Animal Science and Technology，Qingdao Agricutural University，Qingdao 266109，China) |
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| Abstract: | The purified prokaryotic expression of PRV gC functional domain named His-gCN813 was used as immunogen to generate antibodies，and the PRV gC expressed in eukaryotic cell was analysised. The full length of PRV gC was cloned from PRV genome to construct recombinant plasmids p3xFLAG-gC for eukaryotic expression. The expression of PRV gC in p3xFLAG-gC transfected cells and PRV infected cells were analyzed with the anti-gC antibodies by Western blotting and immunofluorescence assay. The results indicated that the eukaryotic expressed system of PRV gC were constructed，gC specific poly-clonal antibodies were generated，detecting 72 ku gC expressed in p3xFLAG-gC transfected cells and 55，72 and 94 ku gC expressed in PRV infected cells，mainly located in the cytoplasm by immunofluorescence. PRV gC would be a potential molecular indicator of PRV，laid foundation for the future research of PRV replication and interaction between PRV and host. |
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| Keywords: | Western blotting |
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