中国药理学通报Issue(11)：1535-1538,4.DOI:10.3969/j.issn.1001-1978.2014.11.011
hi FGF2真核表达载体的构建及其高表达对细胞凋亡的影响
Construction of Hi FGF2 eukaryotic expression plasmids and its over-expression induced cell apoptosis
摘要
Abstract
Aim To construct eukaryotic expressing plasmid of hi FGF2 ( high molecular weight isoform fi-broblast growth factor-2,hi FGF2) gene and to investi-gate its effect on apoptosis after its overexpression in HEK293 cells. Methods The DNA template primer was designed and synthesized. The pDsRed1-N1 plas-mids were digested by the restriction enzymes of Nhel and Hind III. The hi FGF2 was ligated with linearized pDsRed1-N1 by T4 DNA Ligase. The recombinant plasmid was identified by endonuclease digestion and sequenced. The recombinant hi FGF2 plasmid was transient transfected into HEK293 cells by Lipofectami-neTM 2000 Reagent. The transfection efficiency was de-tected by fluorescence inversion microscope. The cell apoptosis was detected by Annexin V-FITC/PI apopto-sis detection kit with flow cytometry analysis. Results The pDsRed1-N1 eukaryotic expression vector was consistent with the design. The recombinant hi FGF2 plasmid was transfected in HEK293 cells. The trans-fection rate was more than 70%. The FITC/PI dyeing rate in hi-FGF2 over-expression HEK297 cells was a-bout ( 29. 12 ± 2. 81 )%. Conclusions pDsRed1-N1 eukaryotic expression vector is successfully constructed and transfected into HEK293 cells. Over-expression of hi FGF2 induces cell apoptosis.关键词
高分子量碱性成纤维细胞生长因子/真核表达载体/转染/细胞凋亡/HEK293细胞/流式细胞术Key words
Hi FGF2/eukaryotic expression vector/transfection/apoptosis/HEK293 cells/flow cytometry分类
医药卫生引用本文复制引用
陈钟琳,姜红岩,洪晓冰,陈中华,郑燕珊,徐涵,石刚刚,黄展勤..hi FGF2真核表达载体的构建及其高表达对细胞凋亡的影响[J].中国药理学通报,2014,(11):1535-1538,4.基金项目
国家自然科学基金资助项目( No 30901810) （ No 30901810）
广东省自然科学基金资助项目(No 9151503102000013) （No 9151503102000013）
