中文摘要：

目的：构建带有组织特异性FLT-1启动子的真核表达载体，检测其在转染的人脐静脉内皮细胞（HUVEC）中对荧光素酶报告基因表达的驱动能力。方法：采用PCR扩增FLT．1启动子，插入到pGL3-Basic-luc载体中，构建携带FLT-1启动子的真核表达载体pGL3-FIJT-Basic-luc，经脂质体法转染HUVEC、HepG2、NIH3T3和HEK293细胞，于转染48h后采用双荧光报告系统检测荧光素酶表达活性。结果：酶切及测序证实构建的pGL3．FLT．Basic-luc载体中含有序列正确的FLT-1基因启动子，双荧光报告系统检测显示，转染的HUVEC细胞其荧光素酶活性明显高于HEK293细胞（P〈0．01），而转染的HepG2和NIH3T3细胞中未检测出荧光素酶表达。结论：克隆的FLT-1启动子具有较高的血管内皮特异性转录活性，可作为血管疾病靶向基因治疗的启动子来源。

英文摘要：

Objective： To construct pGL3 Basic eukaryotic expression vector containing tissues specific promoter of FLT-1 and explore the activity of this promoter in HUVEC cells. Methods ： The FLT-1 gene promoter was amplified by polymerase chain reaction and cloned into pGl_3 Basic vector to construct pGL3 Basic eukaryotic expression vector containing FLT-1 gene promoter（ pGL3-FLT-Basic-luc ）. The purified pGl_3-FLT-Basic-luc was transiently transfected into HUVEC cell and HepG2,NIH3T3,HEK293 cell using liposome transfection reagent, and the activity of luciferase was determined with Dual-Luciferase Reporter Assay System 48h later. Results： DNA sequencing and digestion confirmed that the recombinant of plamid pGI_3-FLT-Basic-luc contained FLT-1 promoter sequence. The activity of luciferase in HUVEC was much higher than in HEK293 after transfection of pGL3-Basic-luc, and little activity of luciferase was detected in other two cells. Conclusion： pGL3 Basic eukaryotic expression vector containing tissues specific promoter of FLT-1 was Successtully constructed, which might be a potential therapeutic reagent for endothelium-targeted gene therapies for vascular disease.