CAJ | 학술논문

目的:构建含有不同片段血管内皮细胞生长因子(VEGF)基因的启动子的萤光素酶报告基因载体,并定位缺氧诱导因子1α(HIF-1α)结合VEGF启动子的区域.方法:以VEGF全长启动子为模板,PCR扩增VEGF启动子的不同片段,插入萤光素酶报告基因载体pGL4-basic,确定所扩增的DNA序列后,将其与HIF-1α表达载体共转染293T细胞,定位HIF-1α结合VEGF启动子的区域.结果:测序结果表明扩增的不同片段的VEGF启动子序列正确;萤光素酶活性实验表明,-1128~-728 bp片段是HIF-1α与VEGF相互作用激活VEGF启动子转录活性的区域.结论:克隆了VEGF启动子5'端缺失突变体报告基因表达载体,为调控VEGF表达的转录因子的筛选及功能研究打下了良好基础.
목적:구건함유불동편단혈관내피세포생장인자(VEGF)기인적계동자적형광소매보고기인재체,병정위결양유도인자1α(HIF-1α)결합VEGF계동자적구역.방법:이VEGF전장계동자위모판,PCR확증VEGF계동자적불동편단,삽입형광소매보고기인재체pGL4-basic,학정소확증적DNA서렬후,장기여HIF-1α표체재체공전염293T세포,정위HIF-1α결합VEGF계동자적구역.결과:측서결과표명확증적불동편단적VEGF계동자서렬정학;형광소매활성실험표명,-1128~-728 bp편단시HIF-1α여VEGF상호작용격활VEGF계동자전록활성적구역.결론:극륭료VEGF계동자5'단결실돌변체보고기인표체재체,위조공VEGF표체적전록인자적사선급공능연구타하료량호기출.
Objective: To clone different regions of vascular endothelial growth factor(VEGF) promoter and insert them into a luciferase reporter gene vector, and to identify the region of VEGF promoter for hypoxia inducible fac?tor-1α(HIF-1α) binding. Methods: Different regions of VEGF promoter were amplified from full length VEGF promoter by PCR and cloned into pGL4-basic. The resulting plasmids were examined by DNA sequencing. Then they were co-transfected with HIF-1α expression plasmid into 293T cell line to analyze promoter activity in the presence of HIF-1α. Results: DNA sequencing showed that the sequences of the promoter regions were correctly cloned. The region -1128~-728 bp of VEGF promoter played a major role in the interaction between HIF-1αand VEGF promoter. Conclusion: The VEGF promoter and 5' terminal deletion mutant reporter genes were cloned successfully, which will be contribute to screen transcription factors that regulate VEGF expression.