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| Identification of tissues specific promoter of FLT-1 and detection its specific activity in transfected HUVEC |
| ZHANG Tao |
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Abstract 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 pGL3 Basic vector to construct pGL3 Basic eukaryotic expression vector containing FLT-1 gene promoter(pGL3-FLT-Basic-luc)．The purified pGL3-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 pGL3-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.
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Received: 18 June 2008
Published: 25 January 2009
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