CAJ | 학술논문

目的研究丙型肝炎病毒非结构蛋白2反式调节基因（NS2TP）的转录调节机制。方法应用PCR方法扩增NS2TP的启动子，克隆入pGL4.10载体，构建萤虫素酶报告基因载体，转染HepG2细胞，检测双萤虫素酶活性，验证其转录活性；通过缺失突变法构建系列截短的NS2TP基因启动子报告基因载体。结果成功构建了系列截短的NS2TP基因启动子报告基因载体，分别命名为N1、N2、N3和N4，并确定了N3为NS2TP基因的核心启动子。结论构建NS2TP基因启动子的系列截短报告基因载体，确定了其最小转录活性区域，为进一步研究NS2TP基因的转录活性调节机制奠定理论基础。
목적연구병형간염병독비결구단백2반식조절기인（NS2TP）적전록조절궤제。방법응용PCR방법확증NS2TP적계동자，극륭입pGL4.10재체，구건형충소매보고기인재체，전염HepG2세포，검측쌍형충소매활성，험증기전록활성；통과결실돌변법구건계렬절단적NS2TP기인계동자보고기인재체。결과성공구건료계렬절단적NS2TP기인계동자보고기인재체，분별명명위N1、N2、N3화N4，병학정료N3위NS2TP기인적핵심계동자。결론구건NS2TP기인계동자적계렬절단보고기인재체，학정료기최소전록활성구역，위진일보연구NS2TP기인적전록활성조절궤제전정이론기출。
Objective To construct series of reporter plasmids with truncated NS2TP gene promoter. Methods Fragments of NS2TP gene promoter was ampliifed by PCR and cloned into pGL4.10-Basic. Then the luciferase reporter vectors of NS2TP gene was constructed. Dual luciferase assays were performed with lysates of HepG2 cells transfected with NS2TP gene promoter reporter plasmids. Results Series of luciferase reporter plasmids with truncated NS2TP gene promoter were successfully constructed and named N1, N2, N3 and N4, respectively. N3 as the minimal promoter was determined. Conclusions Series of luciferase reporter plasmids with truncated NS2TP gene promoter were successfully constructed and their promoter activity were veriifed. These plasmids provide necessary experimental materials for further investigation of regulation of NS2TP gene.