| SARS病毒E蛋白基因与EGFP基因在BHK-21细胞中的融合表达 |
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| 引用本文： | 陶泽新,王志玉,温红玲,任桂杰,宋艳艳,王桂亭,姚苹,许洪芝. SARS病毒E蛋白基因与EGFP基因在BHK-21细胞中的融合表达[J]. 山东大学学报(医学版), 2004, 42(5): 508-510 |
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| 作者姓名： | 陶泽新 王志玉 温红玲 任桂杰 宋艳艳 王桂亭 姚苹 许洪芝 |
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| 作者单位： | 山东大学公共卫生学院病毒学研究室,山东,济南,250012 |
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| 基金项目： | 山东大学SARS研究专项基金资助课题(200304)。 |
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| 摘 要： | 目的：构建SARS病毒E蛋白基因的真核表达载体，观察E蛋白基因在BHK-21细胞中的正常表达及定位。方法：用PCR方法扩增E蛋白基因，上、下游引物两端分别设计EcoRⅠ和BamHⅠ酶切位点，产物双酶切后定向克隆于pEGFP-N1载体，构建真核表达载体pEGFP-E，脂质体法转染BHK-21细胞，荧光显微镜观察E蛋白基因的表达。结果：经酶切和DNA序列测定鉴定，E蛋白基因正确插入到增强绿荧光蛋白EGFP基因上游，荧光显微镜显示E蛋白表达于细胞浆内，包膜上也有分布。结论：在BHK-21细胞中成功表达了SARSCovE-EGFP融合蛋白，蛋白定位于细胞浆内。
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| 关 键 词： | 严重急性呼吸道综合征 E蛋白 基因表达 转染 |
| 文章编号： | 1671-7554(2004)05-0508-03 |
| 修稿时间： | 2004-02-20 |
Expression of SARSCovE-EGFP fused gene in BHK-21 cells |
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| TAO Ze-xin,WANG Zhi-yu,WEN Hong-ling,et al. Expression of SARSCovE-EGFP fused gene in BHK-21 cells[J]. Journal of Shandong University:Health Sciences, 2004, 42(5): 508-510 |
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| Authors: | TAO Ze-xin WANG Zhi-yu WEN Hong-ling et al |
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| Abstract: | Objective: To study the normal expression and location of SARS virus E protein in BHK-21 cells. Methods: The E protein gene was amplified by PCR, and cloned into vector pEGFP-N1, followed by the EGFP(enhanced green fluorescent protein) gene. The recombinant plasmid was transfected into BHK-21 cells with lipofectin and the expression was observed under fluorescence microscope. Results: The two genes of E protein and GFP were fused in codon frame by DNA sequencing. The fluorescence microscopy showed that the E protein was expressed in the cytoplasm and cell membrane of BHK-21 cells. Conclusion: The E protein of SARS virus is expressed successfully together with EGFP. |
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| Keywords: | Severe acute respiratory syndrome Small envelope protein Gene expression Transfection |
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