VEGF165真核表达载体的构建及其在鼠膀胱平滑肌细胞的表达  

Construction of eukaryotic expression vector for VEGF165 gene and its expression in rat bladder smooth muscle cells

作　　者：林茂虎[1] 赵珊[2] 苗芮[3] 贾宁[4] 李娟 

机构地区：[1]第四军医大学研究生院,陕西西安710032 [2]新疆军区联勤部30分部,新疆830011 [3]解放军总医院第一附属医院,北京100037 [4]解放军总医院医院感染与疾病控制科,北京100853 [5]北京二炮通信总站卫生队,北京100085

出　　处：《南方医科大学学报》2007年第5期654-656,659,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(30300349)~~

摘　　要：目的构建血管内皮生长因子(VEGF165)真核表达载体pcDNA3.1(-)/VEGF165,转染鼠膀胱平滑肌细胞并检验其生物学活性。方法将VEGF165cDNA克隆于真核表达载体pcDNA3.1(-),构建真核表达载体pcDNA3.1(-)/VEGF165;并转染入鼠膀胱平滑肌细胞,采用RT-PCR和免疫荧光染色检测VEGF165基因在鼠膀胱平滑肌细胞中的表达,并用MTT法检测转染后细胞上清液中VEGF165的生物学活性。结果和结论将构建的真核表达载体pcDNA3.1(-)/VEGF165转染入鼠膀胱平滑肌细胞后,VEGF的表达增高,转染后细胞上清液具有促使内皮细胞增殖的生物学活性。Objectives To construct an eukaryotic expression vector for vascular endothelial growth factor （VEGF） 165 gene and obtain VEGF expression in rat bladder smooth muscle cells. Methods VEGFI65 eDNA was cloned into the eukaryotic expression vector pcDNA3.1 （-）, and the resultant recombinant vector pcDNA3.1 （-）/VEGF 165 was transfected into the rat bladder smooth muscle cells by electroporation. VEGF expression in the cells was determined by RT-PCR and immunofluorescnce assay, and the biological activity of VEGF in the supemant of the transinfected cell culture was tested by MTT assay. Results and conclusion VEGF expression was obtained in the transinfected cells, and the supemant of the transinfected cell cultures stimulated the proliferation of the endothelial cells.

关 键 词：血管内皮生长因子 真核表达载体 膀胱:平滑肌细胞 

分 类 号：R346[医药卫生—基础医学]