| 猪细小病毒vp2基因的表达和类病毒颗粒的构建 |
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| 引用本文： | 司艳红,方明刚,王汉中.猪细小病毒vp2基因的表达和类病毒颗粒的构建[J].Virologica Sinica,2006,21(2):148-152. |
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| 作者姓名： | 司艳红 方明刚 王汉中 |
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| 作者单位： | 中国科学院武汉病毒研究所分子病毒学重点实验室，中国科学院武汉病毒研究所分子病毒学重点实验室，中国科学院武汉病毒研究所分子病毒学重点实验室 湖北武汉，430071，湖北武汉，430071，湖北武汉，430071 |
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| 基金项目： | 中国科学院二期创新项目资助,项目编号KSCX2-SW-302. |
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| 摘 要： | 利用PCR方法从病毒基因组中获得了vp2完整的编码区并经测序后克隆到原核表达载体pET-32(a)上。在大肠杆菌中诱导表达了VP2与Trx-His-STag的融合蛋白,通过免疫家兔获得了高特异性的兔抗血清。另外,将vp2克隆到pFastBacDUAL载体上,利用昆虫表达系统,即AcMNPV的Bac-to-Bac系统对vp2进行了表达,经SDS-PAGE和Westernblot分析,表达产物为64kD,并且该蛋白能在昆虫细胞中形成类病毒颗粒。
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| 关 键 词： | vp2 猪细小病毒 表达 类病毒颗粒 |
| 收稿时间： | 2005-08-25 |
| 修稿时间： | 2005-10-18 |
Expression of Porcine Parvovirus vp2 Gene and Construction of Virus Like Particles |
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| SI Yan-hong, FANG Ming-gang, WANG Han-zhong.Expression of Porcine Parvovirus vp2 Gene and Construction of Virus Like Particles[J].中国病毒学(英文版),2006,21(2):148-152. |
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| Authors: | SI Yan-hong FANG Ming-gang WANG Han-zhong |
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| Institution: | Key laboratory of Molecular Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071 China |
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| Abstract: | In this study, the open reading frame of PPV vp2 was amplified by PCR, sequenced, cloned and expressed both in prokaryotic and insect cells. In E. coli, by using pET-32(a) expression vector, VP2 was expressed as a Trx-His-S Tag fusion protein with the size of 84 kDa and the protein was used to immunize rabbits for generating specific antibody. Moreover, vp2 gene was cloned into pFastBacDUAL and expressed in baculovirus/insect cell expression system: AcMNPV Bac-to-Bac system. SDS-PAGE and Western-blot analysis showed a single band with the size of 64 kDa. In Sf-21 cells, virus like particles could be observed under electron microscope. |
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| Keywords: | vp2 PPV expression virus like particle |
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