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《China Biotechnology》 2009-01
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Identification of Tissues Specific Promoter of FLT-1 and Detection the Specific Activity in Transfected HUVEC

ZHUGE Fu-yuan1 LV Zhi-mei1 ZHENG Hong-ting2 ZHANG Tao3 DENG Hua-cong1 (1 Department of Endorinology,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)(2 Department of Endorinology,Xinqiao Hospital,Third Military Medical University,Chongqing 400037,China)(3 Key Laboratory of Cell Engineering of Guizhou Province,The Affiliated Hospital of Zunyi Medical College,Zunyi 563003,China)  
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.
【Fund】: 国家自然科学基金资助项目(30570744)
【CateGory Index】: Q343
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