HBV启动子克隆及萤光素酶报告基因载体的构建  

The construction of hepatitis B virus promoter-luciferase reporter gene system

作　　者：余卓[1] 孙学华[1] 李曼[1] 朱晓骏[1] 周振华[1] 乔兵[1] 高月求[1] 

机构地区：[1]上海中医药大学附属曙光医院肝病科,上海市中医临床重点实验室,上海200021

出　　处：《胃肠病学和肝病学杂志》2013年第5期449-451,共3页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家科技重大专项资助项目(No.2012ZX10005-004-002;No.2012ZX10005-010-002-003);国家自然科学基金(No.81102570;No.81072792);国家中医药管理局中医肝胆病重点学科(No.2010sh);上海市自然科学基金(No.10ZR1430900);上海市教育委员会重点学科(第五期)建设资助项目(No.J50307);上海高校创新团队项目(第一期);上海中医药大学优秀团队培养计划;浦东新区中医领军型人才项目;上海市医学领军人才培养计划(No.LJ10004)

摘　　要：目的对HBV启动子基因序列进行分析,克隆并构建S1、S2、ENⅠ/X及ENⅡ/C 4个启动子的萤光素酶报告基因载体。方法用pHBV 1.3作为模板进行PCR扩增目的片段,将获得片段与pGL3-Enhancer荧光素酶报告载体重组构建,分别转染Huh7细胞后,检测萤光素酶活性。结果成功获得121 bp、368 bp、437 bp、237 bp 4个目的扩增片段,并成功构建了S1、S2、ENⅠ/X及ENⅡ/C启动子报告基因载体。结论上述载体的成功构建及初步分析为进一步研究HBV启动子活性及新的抗HBV药物奠定了基础。Objective To analyze the promoter of HBV based on the gene sequence and construct the lueiferase re- port gene vectors with S1, $2, EN I/X and EN Ⅱ/C promoter fragments. Methods The pHBV 1.3 plasmid was used as template for target fragment amplification, and the acquired fragments were constructed with pGL3-Enhancer respec- tively. The luciferase activities were detected after transient transfection of the 4 constructs to Huh7 cell line respective- ly. Results The target fragments with the length of 121 bp, 368 bp, 437 bp and 237 bp were obtained and 4 report gene vectors with S1, S2, EN I/X and EN II/C promoter fragments were constructed successfully. Conclusion These results may make an important basis for the further study of HBV promoter activity and regulation of gene expression, as well as the development of new anti-HBV drugs.

关 键 词：HBV 启动子 pGL3 Enhancer载体 基因克隆 

分 类 号：R575[医药卫生—消化系统]