| 绿色荧光蛋白标记的GRP78蛋白表达和功能研究 |
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| 引用本文： | 钮晓音,李小彦,陈广洁.绿色荧光蛋白标记的GRP78蛋白表达和功能研究[J].南通医学院学报,2009,29(4). |
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| 作者姓名： | 钮晓音 李小彦 陈广洁 |
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| 作者单位： | 上海交通大学医学院,上海免疫学研究所,上海,200025 |
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| 基金项目： | 上海市卫生局青年科研项目资助 |
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| 摘 要： | 目的:应用增强型绿色荧光蛋白(EGFP)来标记葡萄糖调节蛋白78(GRP78),从而研究其与免疫细胞结合的情况。方法:通过在原核表达的GRP78的C末端连接一个EGFP来显示跟踪该蛋白的表达,进而通过流式细胞仪检测该蛋白同小鼠脾脏免疫细胞结合的情况。结果:GRP-EGFP融合蛋白分子量为126kD,Western Blot证实该蛋白表达正确,并且表达GRP78-EGFP的BL21菌在紫外光激发下发射出强烈的绿色荧光;与EGFP单体相比较,GRP78-EGFP主要与中性粒细胞相结合,其平均结合率为5.15%。结论:绿色荧光蛋白标记的GRP78蛋白在原核中的表达具有天然构象并能发挥标记目的蛋白的作用。
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| 关 键 词： | 绿色荧光蛋白 葡萄糖调节蛋白78 原核表达 蛋白标记 |
The study of prokaryotic expression and function of GRP78 tagged with green fluorescence protein |
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| NIU Xiaoyin,LI Xiaoyan,CHEN Guangjie.The study of prokaryotic expression and function of GRP78 tagged with green fluorescence protein[J].ACTA Academiae Medicinae Nantong,2009,29(4). |
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| Authors: | NIU Xiaoyin LI Xiaoyan CHEN Guangjie |
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| Institution: | NIU Xiaoyin; LI Xiaoyan; CHEN Guangjie (Medical School; Shanghai Jiaotong University; Shanghai Institute of Immunology; Shanghai 200025); |
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| Abstract: | Objective:To study the prokaryotic expression and function of GRP78 tagged with green fluorescence protein. Methods:The enhanced green fluorescence protein was linked with the C-terminal end of GRP78 so as to track the expression of GRP78. Moreover,the tagged protein conjugated with splenocytes was determined by FACS. Results:The molecular weight of fusion protein GRP-EGFP was 126 kD and the prokaryotic expression was correctly judged by Western Blot. The bacteria BL21 expressing GRP78-EGFP could emit spark... |
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| Keywords: | Green fluorescence protein Glucose-regulated-protein 78 Prokaryotic expression Protein tagging |
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