医学分子生物学杂志2011，Vol.8Issue(3)：198-203,6.DOI:10.3870/j.issn.1672-8009.2011.03.003
人Tudor-SN基因启动子荧光素酶报告基因表达质粒的构建及活性检测
Construction of pGL3-Basic-Tudor-SN-Promoter-Luciferase Plasmid and Analysis of Its Activity
摘要
Abstract
Objective To construct eukaryotic expressing luciferase recombinant plasmid which contains human Tudor-SN promoter fragment, and to evaluate its activity in HeLa cells. Methods The human Tudor-SN promoter fragments were amplified from genomic DNA extracted from HeLa cells by PCR. and then inserted into pGL3-Basic expression vector at Xho Ⅰ and Hind Ⅲ sites. HeLa cells were cotransfected with the recombinant pGL3-Basic-Tudor-SN-promoter plasmids and β-galactosidase plasmid and detected for luciferase activity after 48 hoursResults Both restriction double enzyme digestion and sequencing analysis confirmed the correct insert of Tudor-SN promoter in the recombinant plasmid pGL3-Basic-Tudor-SN-promoter. The luciferase activity was detected after transfection. Conclusion The recombinant plasmid of pGL3-Basic-Tudor-SN-promoter-luciferase was constructed correctly and expressed effectively, which may be used for further study of regulatory mechanism of Tudor-SN gene in transcription.关键词
人类Tudor-SN蛋白/启动子/重组质粒/虫荧光素酶Key words
human Tudor-SN protein/ promoter/ recombinant plasmid/ luciferase分类
生物科学引用本文复制引用
赵虹,高星杰,葛林,朱梦瑜,段中潮,宋娟,付雪,杨洁..人Tudor-SN基因启动子荧光素酶报告基因表达质粒的构建及活性检测[J].医学分子生物学杂志,2011,8(3):198-203,6.基金项目
国家高技术研究发展计划资助项目(863计划)(No.2007AA02Zll5),国家自然科学基金(No.30670441,30970562,30670802,30811130394,30911130165,30970582),天津市科委国际合作项目(No.07JCZDJC07300),国家自然科学重大研究计划培育项目(No.90919032),国家教育部高等学校博士学科点专项科研基金(No.20091202110001),天津市教委重点项目(No.2008ZD01) （863计划）
