CAJ | 학술논문

目的克隆和鉴定人TSLC1基因的核心启动子区,用于开展其转录调控机制的研究.方法用PCR方法从人基因组DNA中扩增出TSLC1基因翻译起始位点上游一系列大小不等的片段,连入pGL3-Basic荧光素酶报告基因载体中.通过瞬时转染A549及NCI-H446细胞,检测细胞裂解液中的双荧光素酶活性,确定TSLC1基因的核心启动子区.结果在人TSLC1基因启动子区的分段克隆中,ATG上游-68～-329 bp片段在A549及NCI-H446细胞中均具有很强的启动活性,对TSLC1的转录起重要作用.结论人TSLC1基因翻译起始位点ATG上游-68～-329 bp区域可能为基因的核心启动子区.
목적 극륭화감정인TSLC1기인적핵심계동자구,용우개전기전록조공궤제적연구.방법 용PCR방법종인기인조DNA중확증출TSLC1기인번역기시위점상유일계렬대소불등적편단,련입pGL3-Basic형광소매보고기인재체중.통과순시전염A549급NCI-H446세포,검측세포렬해액중적쌍형광소매활성,학정TSLC1기인적핵심계동자구.결과 재인TSLC1기인계동자구적분단극륭중,ATG상유-68～-329 bp편단재A549급NCI-H446세포중균구유흔강적계동활성,대TSLC1적전록기중요작용.결론 인TSLC1기인번역기시위점ATG상유-68～-329 bp구역가능위기인적핵심계동자구.
Objective To clone and to identify the core promoter of human TSLCI used for exploring of transcrip-tion regulatory mechanism.Methods A series of different fragments located in the upstream of translation start site of TSLC1 were amplified from human genomic DNA by PCR,and then constructed into pGL3-Basic luciferase re-porter vector.The activity of different fragments in A549 and NCI-H446 cells was examined by a dual-luciferase as-say after transient transfection,and then the core promoter of TSLC1 was identified.Results Among the different constructs,the fragment of -68 ～ -329 bp located in the upstream of ATG showed the strong activity both in A549 cells and NCI-H446 cells,which played an important role in the transcription of TSLC1.Conclusion The fragment of -68 ～ -329 bp located in the upstream of translation start site of TSLC1 might be the core promoter region.