| 鸡贫血病毒VP1、VP2和VP3基因酵母双杂交载体的构建及其自激活活性的鉴定 |
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| 引用本文： | 孙芬芬,高玉龙,高宏雷,祁小乐,潘伟,王笑梅.鸡贫血病毒VP1、VP2和VP3基因酵母双杂交载体的构建及其自激活活性的鉴定[J].中国兽医科学,2010,40(2). |
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| 作者姓名： | 孙芬芬 高玉龙 高宏雷 祁小乐 潘伟 王笑梅 |
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| 作者单位： | 孙芬芬,高宏雷,祁小乐,潘伟,王笑梅(中国农业科学院,哈尔滨兽医研究所,兽医生物技术国家重点实验室,禽传染病研究室,黑龙江,哈尔滨150001);高玉龙(中国农业科学院,哈尔滨兽医研究所,兽医生物技术国家重点实验室,禽传染病研究室,黑龙江,哈尔滨150001;黑龙江省农业科学院,黑龙江,哈尔滨,150086) |
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| 基金项目： | 现代农业产业技术体系建设专项 |
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| 摘 要： | 为构建鸡贫血病毒(CAV)VP1、VP2、VP3基因酵母双杂交载体并鉴定其自激活作用,从CAV细胞传代毒M9905毒株中提取基因组DNA,利用PCR分别扩增VP1、VP2和VP3基因,采用Gateway技术将VP1、VP2和VP3基因分别克隆到酵母双杂交载体pDEST32和pDEST22中,构建了诱饵载体及猎物载体,经酶切和PCR鉴定且测序正确后,用醋酸锂法转化酵母菌株Mav203,通过缺陷型平板SD/-Leu/-Trp/-His和SD/-Leu/-Trp/-Ura筛选以及LacZ报告基因检测载体有无自激活作用。结果表明,成功构建了诱饵载体pDEST32-VP1/VP2/VP3和捕获载体pDEST22-VP1/VP2/VP3,并证明其VP1、VP2和VP3蛋白在酵母双杂交系统中无自激活作用。研究结果为下一步利用酵母双杂交系统探索CAVVP1、VP2、VP3蛋白之间的相互作用奠定了基础。
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| 关 键 词： | 鸡贫血病毒 蛋白相互作用 酵母双杂交载体 自激活作用 |
Construction of bait vectors with VP1,VP2 and VP3 genes of CAV and identification of its self-activation in yeast two-hybrid system |
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| SUN Fen-fen,GAO Yu-long,GAO Hong-lei,QI Xiao-le,PAN Wei,WANG Xiao-mei.Construction of bait vectors with VP1,VP2 and VP3 genes of CAV and identification of its self-activation in yeast two-hybrid system[J].Veterinary Science in China,2010,40(2). |
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| Authors: | SUN Fen-fen GAO Yu-long GAO Hong-lei QI Xiao-le PAN Wei WANG Xiao-mei |
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| Institution: | SUN Fen-fen1,GAO Yu-long1,2,GAO Hong-lei1,QI Xiao-le1,PAN Wei1,WANG Xiao-mei1(1.State Key Laboratory of Veterinary Biotechnology/Division of Avian Infectious Diseases,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China,2.Heilongjiang Academy of Agricultural Sciences,Harbin 150086,China) |
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| Abstract: | To construct the yeast two-hybrid bait vectors with VP1,VP2 and VP3 genes of chicken anemia virus(CAV) and identify their self-activations,the VP1,VP2 and VP3 genes were amplified by PCR from the CAV M9905 strain total genomic DNA and then cloned them into pDEST32 and pDEST22 vectors using Gateway technology,respectively.The restriction enzyme digestion,PCR and sequence analysis were performed to confirm the fact that those genes had been inserted successfully into the vectors,then pDEST32-VP1/VP2/VP3,pDEST... |
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| Keywords: | chicken anemia virus protein interaction yeast two-hybrid vector self-activation |
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