Expression of Porcine Parvovirus vp2 Gene and Construction of Virus Like Particles

SI Yan-hong; FANG Ming-gang; WANG Han-zhong

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April 2006

SI Yan-hong, FANG Ming-gang and WANG Han-zhong. Expression of Porcine Parvovirus vp2 Gene and Construction of Virus Like Particles[J]. Virologica Sinica, 2006, 21(2): 148-152.

Citation: SI Yan-hong, FANG Ming-gang, WANG Han-zhong. Expression of Porcine Parvovirus vp2 Gene and Construction of Virus Like Particles .VIROLOGICA SINICA, 2006, 21(2) : 148-152.

猪细小病毒vp2基因的表达和类病毒颗粒的构建

1.
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

通讯作者： 王汉中,

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-S Tag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Western blot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。
- vp2
- , 猪细小病毒
- , 表达
- , 类病毒颗粒

Expression of Porcine Parvovirus vp2 Gene and Construction of Virus Like Particles

1.
Key laboratory of Molecular Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China

Corresponding author: WANG Han-zhong,

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.
- vp2
- , PPV
- , expression
- , virus like particle

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Expression of Porcine Parvovirus vp2 Gene and Construction of Virus Like Particles

Corresponding author: WANG Han-zhong,

1. Key laboratory of Molecular Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China

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.