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。
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...
出处
《南通大学学报（医学版）》
2009年第4期235-238,241,共5页
Journal of Nantong University(Medical sciences)
基金
上海市卫生局青年科研项目资助(2008Y019)
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
Green fluorescence protein
Glucose-regulated-protein 78
Prokaryotic expression
Protein tagging
