CAJ | 학술논문

目的：探讨乙型肝炎病毒（HBV）前S2蛋白（preS2）对人酰基蛋白硫酯酶1（APT1）启动子的作用。方法生物信息学方法确定人APT1启动子序列。PCR扩增人APT1启动子和HBV preS2基因，分别插入pGL3和pcDNA3．1（－）质粒构建人APT1启动子荧光素酶报告基因质粒 pGL3‐APT1和 HBV preS2真核表达质粒 pcDNA3．1（－）‐preS2。将 pGL3‐APT1和pcDNA3．1（－）‐preS2共转染人肝癌细胞系 HepG2，然后通过检测细胞荧光素酶活性来评价 preS2对人APT1基因启动子的作用。数据用独立样本 t检验分析。结果测序结果证实pcDNA3．1（－）‐preS2与pGL3‐APT1与实验设计相符。pGL3‐APT1的荧光素酶活性是阳性对照质粒pGL3‐Control的荧光素酶活性的1．2倍（P＜0．01）。pcDNA3．1（－）‐preS2与pGL3‐APT1共转染HepG2细胞的荧光素酶活性为无preS2基因质粒pcDNA3．1（－）与pGL3‐APT1共转染 HepG2细胞荧光素酶活性的2．6倍（P＜0．01）。结论本研究克隆的人APT1启动子序列具有高启动子活性；HBV preS2可激活人APT1启动子。
목적：탐토을형간염병독（HBV）전S2단백（preS2）대인선기단백류지매1（APT1）계동자적작용。방법생물신식학방법학정인APT1계동자서렬。PCR확증인APT1계동자화HBV preS2기인，분별삽입pGL3화pcDNA3．1（－）질립구건인APT1계동자형광소매보고기인질립 pGL3‐APT1화 HBV preS2진핵표체질립 pcDNA3．1（－）‐preS2。장 pGL3‐APT1화pcDNA3．1（－）‐preS2공전염인간암세포계 HepG2，연후통과검측세포형광소매활성래평개 preS2대인APT1기인계동자적작용。수거용독립양본 t검험분석。결과측서결과증실pcDNA3．1（－）‐preS2여pGL3‐APT1여실험설계상부。pGL3‐APT1적형광소매활성시양성대조질립pGL3‐Control적형광소매활성적1．2배（P＜0．01）。pcDNA3．1（－）‐preS2여pGL3‐APT1공전염HepG2세포적형광소매활성위무preS2기인질립pcDNA3．1（－）여pGL3‐APT1공전염 HepG2세포형광소매활성적2．6배（P＜0．01）。결론본연구극륭적인APT1계동자서렬구유고계동자활성；HBV preS2가격활인APT1계동자。
Objective To investigate the trans‐regulative effect of hepatitis B virus (HBV) preS2 on the promoter of human acyl protein thioesterase 1 (APT1) gene .Methods The promoter sequence of human APT1 gene was identified applying the soft‐ware of bioinformatics .The APT1 promoter and HBV preS2 gene were amplified with PCR and cloned into pGL3 and pcDNA3 .1 (-) plasmids to construct the luciferase reporter gene plasmid of human APT1 gene promoter pGL3‐APT1 and the preS2 eukary‐otic expression plasmid pcDNA3 .1(-)‐preS2 ,respectively .The effect of the preS2 on the human APT1 gene promoter was exam‐ined by cotransfecting hepatocellular carcinoma cell HepG2 with pGL3‐APT1 and pcDNA3 .1(-)‐preS2 and measuring luciferase activities of the HepG2 cells .The statistical data were analyzed with independent‐samples t test .Results Both plasmids of pGL3‐APT1 and pcDNA3 .1(-)‐preS2 were confirmed by DNA sequencing to be accurately constructed as design .The luciferase activity of the pGL3‐APT1 was 1 .2 times (P<0 .01) that of the positive control plasmid pGL3‐Control .And the luciferase activity of the HepG2 cells cotransfected with pcDNA3 .1(-)‐preS2 and pGL3‐APT1 was 2 .6 times (P<0 .01) that of the HepG2 cells cotrans‐fected with the plasmid without preS2 gene pcDNA3 .1(-) and pGL3‐APT1 .Conclusion The human APT1 promoter cloned in the study has high promoter activity ;HBV preS2 activates human APT1 promoter .