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| CD24在人胚肾上皮细胞Hek293中的表达及检测 | |
| 引用本文： | 敖翔,章清,武强,贺乐奇,张群峰,柯文才,彭志海. CD24在人胚肾上皮细胞Hek293中的表达及检测[J]. 中国现代医生, 2012, 0(29): 47-48,51 |
| 作者姓名： | 敖翔 章清 武强 贺乐奇 张群峰 柯文才 彭志海 |
| 作者单位： | [1]复旦大学附属上海市第五人民医院检验科,上海200040 [2]上海交通大学附属第一人民医院普外科,上海200030 |
| 基金项目： | 国家自然科学基金（81072008） |
| 摘 要： | 目的构建含有CD24基因的重组质粒,在人胚肾上皮细胞Hek293中表达出具有免疫原性的抗原。方法用PCR方法扩增CD24基因,克隆到载体pYD5-hFC中,获得重组质粒pYD5-CD24,转化DH10α感受态细胞中,挑选克隆抽取质粒,以脂质体介导将质粒转染Hek293细胞,72 h后收集细胞培养上清,Western-Blot分析鉴定。结果构建了含有CD24基因的重组质粒,并在人胚肾上皮细胞Hek293中获得带有hFC标签的CD24蛋白。结论本研究成功表达出了具有免疫原性的CD24蛋白。 |
| 关 键 词： | CD24 表达 Western |
Expression of CD24 gene in human Hek293 cells |
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| AO Xiang,HANG Qing,WU Qiang,HE Leqi,ZHANG Qunfeng,KE Wencai,PENG Zhihai. Expression of CD24 gene in human Hek293 cells[J]. , 2012, 0(29): 47-48,51 | |
| Authors: | AO Xiang HANG Qing WU Qiang HE Leqi ZHANG Qunfeng KE Wencai PENG Zhihai |
| Affiliation: | (l.Department of Clinical Laboratory,Fudan University Affilicated Fifth People＇s Hospital of Shanghai City, Shanghai 200040,China; 2.Department of General Surgery,Shanghai Jiaotong University Affiliated First People＇s Hospital,Shanghai 200030, China) |
| Abstract: | Objective To construct the recombinant plasmid with gene CD24 and to express immunogenic protein in human embryo kidney Hek293 cells. Methods CD24 gene was cloned into the vector pYD5-hFC by PRC method. The required recombinant plasmids pYD5-CD24 transformed into the GH10α component cells. Then the selected recombinant plasmids were transfected with lipofectamine reagent into Hek293 cells. After 72 hours,the protein in the culture supernatant of the Hek293 cells were collected and analyzed by western blot. Results The recombinant plasmid transfected with CD24 gene were constructed. It was detected out that the CD24 protein with hFC sign were expressed in human embryo kidney Hek293 cells. Conclusion The CD24 gene expressed successfully in the transfected Hek293 cells and had good immunogenic activity. |
| Keywords: | CD24 Expression Western |
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