| 鸽圆环病毒浙江株△Cap基因的克隆与原核表达 |
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| 引用本文： | 余旭平,刘晓宁,郑新添,邬栋栋,刘江梅,竺春.鸽圆环病毒浙江株△Cap基因的克隆与原核表达[J].中国预防兽医学报,2007,29(9):680-684. |
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| 作者姓名： | 余旭平 刘晓宁 郑新添 邬栋栋 刘江梅 竺春 |
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| 作者单位： | 1. 浙江大学,动物科学学院,浙江,杭州,310029
2. 龙岩学院,生物科学与工程系,浙江,龙岩,364000 |
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| 摘 要： | 根据已发表的鸽圆环病毒(PiCV)序列,在V1ORF保守序列部位设计引物(目的片段长度为629bp),对浙江某鸽养殖场的病料进行PCR扩增检测,获得阳性样品;应用设计的删除核定位信号外壳蛋白基因(△Cap)的引物,扩增获得680bp的△Cap基因,克隆于pMD18-T载体,进行测序;将△Cap基因亚克隆到原核表达载体pET-28a进行融合表达,SDS-PAGE电泳检测发现,△Cap融合蛋白经IPTG诱导后在大肠杆菌中以包涵体形式表达,目的蛋白表达量占菌体总蛋白的22.1%。
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| 关 键 词： | 鸽 圆环病毒 核定位信号 △Cap蛋白 表达 |
| 文章编号： | 1008-0589（2007）09-0680-05 |
| 修稿时间： | 2007-03-12 |
Cloning and prokaryotic expression of the △Cap gene of pigeon circovirus isolated from Zhejiang province |
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| YU Xu-ping,LIU Xiao-ning,ZHENG Xin-tian,WU Dong-dong,LIU Jiang-mei,ZHU Chun.Cloning and prokaryotic expression of the △Cap gene of pigeon circovirus isolated from Zhejiang province[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(9):680-684. |
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| Authors: | YU Xu-ping LIU Xiao-ning ZHENG Xin-tian WU Dong-dong LIU Jiang-mei ZHU Chun |
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| Institution: | 1. Aniamal Science College, Zhejiang University, Hangzhou 310029, China; 2. Bioscience and Bioengineering Department, Longyan College, Longyan 364000, China |
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| Abstract: | A DNA fragment of V1 ORF was amplified from samples in a pigeon farm in Zhejiang Province using primers designed based on the published pigeon circovirus (PiCV) sequences. A truncated Cap gene without the coding region of nuclear localization signal was PCR amplified, cloned into pMD18-T and sequenced. The gene was then subcloned into the pET-28a and transferred into E.coli for expression. SDS-PAGE analysis showed that the recombinant protein was expressed in the form of inclusion body and accumulated up to 22.1 % of the total bacteria protein. |
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