汉坦病毒GM04-38株包膜糖蛋白基因表达载体的构建及表达

山东大学学报(医学版)

汉坦病毒GM04-38株包膜糖蛋白基因表达载体的构建及表达

王炳花¹,陶泽新³,王丽娟¹,曹海霞¹,王志玉^1,2

1. 山东大学公共卫生学院病毒学研究室, 济南 250012; 2. 实验畸形学教育部重点实验室, 济南 250012;3. 山东省疾病预防控制中心, 济南 250014

收稿日期:2007-11-05 修回日期:1900-01-01 出版日期:2008-01-16 发布日期:2008-01-16
通讯作者: 王志玉

Construction and expression of expressional vectors of envelope glycoprotein genes of hantavirus, GM04-38 strain

WANG ping-hua¹,TAO Ze-xin³,WANG Li-juan¹,CAO Hai-xia¹,WANG Zhi-yu^1,2

1. Department of Virology, School of Public Health, Shandong University;2. Key Laboratory of Experimental Teratology, Ministry of Education;3. Shandong Center for Disease Control and Prevention

Received:2007-11-05 Revised:1900-01-01 Online:2008-01-16 Published:2008-01-16
Contact: WANG Zhi-yu

摘要/Abstract

摘要： 摘要：目的构建汉坦病毒GM04-38株包膜糖蛋白G1、G2的真核表达载体并将其在Vero E6细胞中进行表达,观察细胞融合现象。方法采用PCR和基因克隆技术,将GM0438株G1、G2基因分别克隆到表达载体pCAGGS／MCS,酶切和测序鉴定正确后,将表达载体pCAGGS-G1、pCAGGS-G2共转染Vero E6细胞,酸性MEM处理后观察细胞融合现象的产生,并以间接免疫荧光试验(IFA)观察糖蛋白的表达情况。结果显示在Vero E6细胞中成功表达出糖蛋白G1、G2,IFA显示荧光信号分布在细胞浆中。二者的共表达可产生良好的生物学活性,在偏酸性条件下引起Vero E6细胞发生融合,而转染pCAGGS-NP的细胞没有融合现象发生。共表达也未出现明显的促进效应。结论成功构建了糖蛋白G1、G2的表达载体，并在Vero E6细胞中呈有效表达,二者的共表达可在偏酸性条件下引起Vero E6细胞发生融合。

Abstract: To construct expressional vector of glycoprotein G1 and G2 of Hantavirus, GM04-38 strain, express it in Vero E6 cells, and to observe cell fusions. MethodsBy PCR and the gene cloning method, Hantavirus GM0438 strain G1 and G2 genes were cloned into plasmid pCAGGS／MCS. After being confirmed by enzyme digestion and sequence analysis, the pCAGGS-G1 and pCAGGS-G2 were co-transfected into Vero E6 cells. After being treated with acidic MEM, the cells were fixed and stained by Giemsa and were observed under microscopy. IFA was also used to observe the expression efficiencies. ResultsIFA showed the G1 and G2 genes were successfully expressed, and the fluorescent signal was robust and concentrated in the perinuclear region of the transfected cells. Giemsa staining showed that cellcell fusion could be induced under the co-expression of G1 and G2 under acidic conditions, while the coexpression of the NP gene could not enhance the fusion. ConclusionThe expressional vectors of glycoprotein G1 and G2 are successfully constructed and expressed in Vero E6 cells,

Key words: Hantavirus, Glycoprotein, Expressional vector, Cell fusion

中图分类号:

R373.32

王炳花,陶泽新,王丽娟,曹海霞,王志玉,. 汉坦病毒GM04-38株包膜糖蛋白基因表达载体的构建及表达[J]. 山东大学学报(医学版), 2008, 46(1): 68-71.

WANG ping-hua,TAO Ze-xin,WANG Li-juan,CAO Hai-xia,WANG Zhi-yu,. Construction and expression of expressional vectors of envelope glycoprotein genes of hantavirus, GM04-38 strain[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2008, 46(1): 68-71.

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