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.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
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)~~
关键词
血管内皮生长因子
真核表达载体
膀胱:平滑肌细胞
vascular endothelial growth factor
eukaryotic expression vector
bladder
smooth muscle cells
