Expression of Helicobacter pylori cagA gene in Insect Cells by Baculovirus

LIANG Jun; GONG Min; YUAN Zhi—ming’’; LIANG Bu—feng

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August 2002

LIANG Jun, GONG Min, YUAN Zhi—ming’’ and LIANG Bu—feng. Expression of Helicobacter pylori cagA gene in Insect Cells by Baculovirus[J]. Virologica Sinica, 2002, 17(4): 336-339.

Citation: LIANG Jun, GONG Min, YUAN Zhi—ming’’, LIANG Bu—feng. Expression of Helicobacter pylori cagA gene in Insect Cells by Baculovirus .VIROLOGICA SINICA, 2002, 17(4) : 336-339.

利用杆状病毒表达幽门螺杆菌cagA 基因

1.
中国科学院武汉病毒研究所．湖北武汉430071

摘要

幽门螺杆菌cag A 基因克隆到杆状病毒表达系统的pBlueBacHis2A转移载体中，将重组质粒pBlueBacHis2A。 CagA与亲本病毒Bac．N．blue DNA共转染Sf9细胞，以空斑法纯化获得的重组杆状病毒。经PCR法鉴定后进行扩增培养，SDS-PAGE和Western bolt检测结果证实所表达的蛋白为CagA蛋白，间接ELISA分析表明。表达产物可与 Hp感染者血清发生特异性的免疫反应。
- 幽门螺杆菌
- , cagA基因
- , 表达
- , 昆虫细胞

Expression of Helicobacter pylori cagA gene in Insect Cells by Baculovirus

1.
Wuhan Institute of virology， The Chinese Academy of Sciences，Wuban 430071，China

Abstract

A recombination transposing vector pBlueBacHis2一CagA was constructed by inserting Heli— cobacter pylori f口gA gene into pBlueBacHis2A vector of baculovirus expession system ．Co—transfecting Sf9 cells with Bac—N—blue DNA and pBlueBacHis2一CagA，recombinant viral DNA was obtained by pu— rification of recombinant plaques．PCR result showed ca gene intergrating in correct position of bac— ulovirus gemone．The expression of Helicobacter pylori ca gene was confirmed by SDS-PAGE and W esternbolt．Using indirect ELISA assay，it was found that the protein exhibited good specific reac— tivity with sera of HP—infected individuals
- Helicobacter pylori
- , cagA gene
- , Expression
- , Insect Cell

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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 Helicobacter pylori cagA gene in Insect Cells by Baculovirus

1. Wuhan Institute of virology， The Chinese Academy of Sciences，Wuban 430071，China

Abstract: A recombination transposing vector pBlueBacHis2一CagA was constructed by inserting Heli— cobacter pylori f口gA gene into pBlueBacHis2A vector of baculovirus expession system ．Co—transfecting Sf9 cells with Bac—N—blue DNA and pBlueBacHis2一CagA，recombinant viral DNA was obtained by pu— rification of recombinant plaques．PCR result showed ca gene intergrating in correct position of bac— ulovirus gemone．The expression of Helicobacter pylori ca gene was confirmed by SDS-PAGE and W esternbolt．Using indirect ELISA assay，it was found that the protein exhibited good specific reac— tivity with sera of HP—infected individuals