摘要
目的 研究血管内皮生长因子基因 (VEGF12 1)在原代培养大鼠肝细胞中转染表达。方法 构建 pIRES EYFP /VEGF12 1质粒 ,逆转录 聚合酶链反应 (RT PCR)、WesternBlot和荧光显微镜观察VEGF基因在原代培养大鼠肝细胞中的转染表达。结果 pIRES EYFP/VEGF12 1质粒成功构建并转染原代培养大鼠肝细胞 ,RT PCR观察到 2 5 8bp的VEGF特异性条带 ,WesternBlot检测到相对分子质量 42× 10 3 的特异性条带 ,荧光显微镜观察到转染阳性肝细胞发出黄绿色荧光。结论 pIRES EYFP/VEGF12 1质粒在原代培养大鼠肝细胞转染并表达 ,为VEGF基因修饰肝细胞移植和基因治疗奠定基础。
Objective To investigate the transfection and expression of vascular endothelial growth factor genes (VEGF 121) in primary cultured rat hepatocyte.Methods After construction of vector pIRES-EYFP /VEGF 121,the transfection and expression of exogenous VEGF 121 gene in primary cultured rat hepatocyte was observed by reverse transcriptase polymerase chain reaction (RT-PCR),Western Blotb and fluorescent microscopy.Results pIRES-EYFP/VEGF 121 plasmid was constructed and transfected into primary cultured rat hepatocytes successfully.A special blot about 258bp was observed by RT-PCR,an about 42×103 special blot was examined by Western blotb,and yellow-green fluorescence was found under a fluorescent microscope.Conclusion pIRES-EYFP/VEGF 121 plasmid was transfected and expressed successfully in primary cultured rat hepatocyte,which may contribute to hepatocyte transplantation and gene therapy after hepatocyte modificated by VEGF gene.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2003年第3期236-237,T002,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目 (39670 71 5)
国家教委博士点基金资助项目 (9947)
