摘要
目的 :以质粒 PCNDA3.0为载体 ,研究血管内皮生长因子 ( VEGF)目的基因在体外真核细胞中的表达。方法 :将编码人可溶性 VEGF165的目的基因克隆于带有 CMV启动子 /增强子的PCNDA3.0载体上 ,重组为 PCNDA3.0 - VEGF165质粒载体。直接以质粒 DNA形式应用脂质体介导的方法转染人脐静脉内皮细胞 ECV30 4 ,收取细胞上清 ,用 ELISA和 Western blot方法检测上清中 VEGF165可溶性表达产物。结果 :VEGF165基因在 CMV启动子 /增强子的调控下 ,可在ECV30 4细胞高效表达 ,且在转染后 72 h表达量明显高于转染后 48h的表达。结论 :以质粒PCNDA3.0为载体 ,VEGF165基因可以在体外真核细胞中表达 。
Objective: To construct a plasmid vector encoding human vascular endothelial growth factor (VEGF) and analyse its expression in eukaryotic cells in vitro. Methods: A soluble human VEGF 165 gene was cloned into the vector PCNDA3.0 which contained the human cytomegalovirus (CMV) promotor/enhancer. The recombined PCNDA3.0 VEGF vector was used directly to transfect to endothelial cells of the human umbilical vein ECV304. The cells were transfected with liposome. The supernatant then was collected and the soluble protein of VEGF gene expression in the supernatant was analysed with ELISA and Western blot.Results: The recombined plasmid vector might express VEGF in ECV304 cell line under the regulation of human CMV promotor/enhancer. Conclusion: This vector with PCNDA3.0 VEGF can express VEGF in eukaryotic cells in vitro and may be useful in the treatmant of ischemic disorders.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2002年第1期11-13,共3页
Journal of Jilin University：Medicine Edition
关键词
血管内皮生长因子
基因表达
质粒
重组DNA
vascular endothelial growth factor (VEGF)
gene expression
recombinant DNA
