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Establishment of a Cell-Based Assay to Screen Regulators of the Hypoxia-Inducible Factor-1-Dependent Vascular Endothelial Growth Factor Promoter
Ji, Deng Bo1; Zhu, Hai Bo3,4; Ye, Jia1; Li, Chang Ling2
关键词vascular endothelial growth factor hypoxia-inducible factor-1 cell-based assay hypoxia response element luciferase reporter gene
刊名BIOLOGICAL & PHARMACEUTICAL BULLETIN
2008-12-01
31期:12页:2255-2259
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Pharmacology & Pharmacy
资助者National Natural Sciences Foundation of China ; National Natural Sciences Foundation of China
研究领域[WOS]Pharmacology & Pharmacy
关键词[WOS]IN-VITRO ; ANGIOGENESIS ; TRANSCRIPTION
英文摘要

In this study, we established a drug screening system based on transcriptional regulation of vascular endothelial growth factor (VEGF) under hypoxia-inducible factor-1 alpha a control. We cloned the neomycin-resistance gene into the plasmid GL (pGL)3-promoter vector to generate the pGL3-promoter-neo vector. The 3 copies of the 47-bp fragment that contained the hypoxia response element of VEGF were synthesized and inserted in front of the minimal promoter of the pGL3-promoter-neo vector to generate p3HRE-luc-neo. The recombinant reporter gene vectors were transfected into EAhy926 cells, and stable cell lines were obtained. The positive cell line was selected for its ability to express luciferase in response to hypoxia. We demonstrated that CoCl, significantly enhances luciferase activity in a concentration-dependent fashion. We then optimized the cell density and incubation time under hypoxia which were used to screen. The assay exhibited a low background and was an ideal model for high-throughput screening for human VEGF regulators.

语种英语
所属项目编号30370720 ; 30572343
资助者National Natural Sciences Foundation of China ; National Natural Sciences Foundation of China
WOS记录号WOS:000261204600015
引用统计
被引频次:3[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/58989
专题北京大学药学院
作者单位1.Peking Univ, Sch Pharmaceut Sci, Dept Mol & Cellular Pharmacol, Beijing 100191, Peoples R China
2.Peking Univ, Sch Pharmaceut Sci, State Key Lab Nat & Biomimet Drugs, Beijing 100191, Peoples R China
3.Chinese Acad Med Sci, Chinese Herbal Med Affiliated Minist Educ, Inst Mat Med, Key Lab Bioact Subst & Resources Utilizat, Beijing 100050, Peoples R China
4.Peking Union Med Coll, Beijing 100050, Peoples R China
推荐引用方式
GB/T 7714
Ji, Deng Bo,Zhu, Hai Bo,Ye, Jia,et al. Establishment of a Cell-Based Assay to Screen Regulators of the Hypoxia-Inducible Factor-1-Dependent Vascular Endothelial Growth Factor Promoter[J]. BIOLOGICAL & PHARMACEUTICAL BULLETIN,2008,31(12):2255-2259.
APA Ji, Deng Bo,Zhu, Hai Bo,Ye, Jia,&Li, Chang Ling.(2008).Establishment of a Cell-Based Assay to Screen Regulators of the Hypoxia-Inducible Factor-1-Dependent Vascular Endothelial Growth Factor Promoter.BIOLOGICAL & PHARMACEUTICAL BULLETIN,31(12),2255-2259.
MLA Ji, Deng Bo,et al."Establishment of a Cell-Based Assay to Screen Regulators of the Hypoxia-Inducible Factor-1-Dependent Vascular Endothelial Growth Factor Promoter".BIOLOGICAL & PHARMACEUTICAL BULLETIN 31.12(2008):2255-2259.
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