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| HMGB1启动子红色荧光蛋白报告基因载体的构建及在机械牵张诱导下的活性检测 | |
| 引用本文： | 丁宁,肖慧,高巨,许立新,余守章. HMGB1启动子红色荧光蛋白报告基因载体的构建及在机械牵张诱导下的活性检测[J]. 第三军医大学学报, 2009, 31(22): 2237-2239 |
| 作者姓名： | 丁宁 肖慧 高巨 许立新 余守章 |
| 作者单位： | 1. 广州市第一人民医院麻醉科和麻醉学实验室,广州,510180 2. 广州市第一人民医院门诊部,广州,510180 3. 广州中医药大学第二附属医院麻醉科,广州,510120 |
| 基金项目： | 广州市医药卫生科技一般引导项目，重点项目，广东省医学科学研究基金，广东省科技计划项目 |
| 摘 要： | 目的 构建高迁移率族蛋白B1(high mobility group box 1 protein,HMGB1)启动子的红色荧光蛋白报告基因载体.方法 采用基因重组技术构建含HMGB1启动子区的红色荧光蛋白报告基因载体pDsRedl-1-HMGB1P.经PCR、酶切和DNA测序鉴定后,将重组载体转染HEK293细胞,在荧光显微镜下观察其在细胞内的表达以及对机械牵张刺激的反应.以转染空载体pDsRedl-1的细胞作为对照.结果 PCR、酶切和DNA测序表明重组载体pDsRedl-1-HMGB1P的构建正确,该表达载体在HEK293细胞静息状态下表达水平较低,经机械牵张刺激后,在荧光显微镜下可看到高亮度的红色荧光.转染空载体pDsRedl-1的细胞未见红色荧光.结论 成功构建了HMGB1启动子的红色荧光蛋白报告基因载体,对机械牵张刺激有很好的反应. |
| 关 键 词： | 高迁移率族蛋白B1 红色荧光蛋白 基因 报告 机械牵张 |
Construction of red fluorescent protein reporter gene vector containing HMGB1 promotor and its activity under mechanical stretch |
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| DING Ning,XIAO Hui,GAO Ju,XU Li-xin,SHE Shou-zhang. Construction of red fluorescent protein reporter gene vector containing HMGB1 promotor and its activity under mechanical stretch[J]. Acta Academiae Medicinae Militaris Tertiae, 2009, 31(22): 2237-2239 | |
| Authors: | DING Ning XIAO Hui GAO Ju XU Li-xin SHE Shou-zhang |
| Affiliation: | DING Ning1,XIAO Hui2,GAO Ju3,XU Li-xin1,SHE Shou-zhang1(1Department of Anesthesiology,2Department of Outpatient,First People's Hospital of Guangzhou,Guangzhou 510180,3Department of Anesthesiology,Second Affiliated Hospital,Guangzhou University of Traditional Chinese Medicine,Guangzhou 510120,China) |
| Abstract: | Objective To construct the red fluorescent protein reporter gene vector containing high mobility group box 1 protein(HMGB1) promoter sequence and study the regulation mechanism of the expression of HMGB1gene under mechanical stretch.Methods HMGB1 promoter was subcloned into a red fluorescent protein vector,pDsRed1-1.After identified by PCR,enzyme digestion and DNA sequencing,the recombinant vector pDsRed1-1-HMGB1P was then transfected into HEK293 cells.Blank vector or pDsRed-1 was transfected into 293 cells... |
| Keywords: | high mobility group box 1 protein red fluorescent protein reporter gene mechanical stretch |
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