[1]马云云),冯龙),卫艳萍),等.带新霉素抗性的萤光素酶报告载体的构建*[J].郑州大学学报(医学版),2009,(03):533-536.
 MA Yunyun),FENG Long),WEI Yanping),et al.Construction of a luciferase reporter vector with resistance of neomycin[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(03):533-536.
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带新霉素抗性的萤光素酶报告载体的构建*()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2009年03期
页码:
533-536
栏目:
论著
出版日期:
2009-05-01

文章信息/Info

Title:
Construction of a luciferase reporter vector with resistance of neomycin
作者:
马云云1)冯龙1)卫艳萍1)李月白2)何婷玉1)董子明3)赵国强1)#
1)郑州大学基础医学院微生物学与免疫学教研室郑州4500012)郑州大学基础医学院生物化学教研室郑州4500013)郑州大学基础医学院病理生理学教研室郑州450001
Author(s):
MA Yunyun1)FENG Long1)WEI Yanping1)LI Yuebai2)HE Tingyu1)etc
关键词:
萤光素酶报告载体pGL3新霉素抗性基因neo克隆
Keywords:
luciferase reporter vectorneomycin resistance gene neoclone
分类号:
R965.1
摘要:
3neoCMV promoter稳定转染组细胞萤光素酶活性。结果:PCR扩增出neo表达单元;酶切和DNA测序分析鉴定得到带新霉素抗性的萤光素酶报告质粒pGL3neo。G418筛选可获得稳定转染pGL3neo细胞株和稳定转染pGL3neoCMV promoter的细胞株。荧光检测显示:瞬时转染pGL3组、pGL3CMV promoter组及稳定转染pGL3neo组、pGL3neoCMV promoter组萤光素酶活性值分别为1 047±86、1 504±107和34 819±1 479、456 109±15 791,稳定转染组萤光素酶活性均高于瞬时转染组,P<0.05。结论:成功构建了带新霉素抗性的萤光素酶报告基因载体,使报告基因检测数据的敏感性和可靠性显著提高。
Abstract:
To construct a luciferase reporter vector with neomycin resistance.Methods:To design and amplify neomycin resistance gene, then clone the gene into luciferase reporter vector pGL3,and obtain the luciferase reporter vector pGL3neo.To compare the luciferase activity of cell lines transiently transfected by pGL3 and pGL3CMV promoter, and cell line lasting transfected by pGL3neo and pGL3neoCMV promoter after screening.

参考文献/References:

[1]Cheng L, Li L, Qiao X, et al. Functional characterization of the promoter of human kinetochore protein HEC1: novel link between regulation of the cell cycle protein and CREB family transcription factors[J]. Biochim Biophys Acta,2007,1 769(9/10):593
[2]HerrmannLavoie C, Rao CV, Akoum A. Chorionic gonadotropin downregulates the expression of the decoy inhibitory interleukin 1 receptor type II in human endometrial epithelial cells[J]. Endocrinology,2007,148(11):5 377
[3]高虹,欧阳克清,郑旭煦,等. 毒蕈胆碱M1受体激动剂的高通量筛选模型[J]. 中国药理学通报, 2003,19(7):776
[4]唐建华,高小平,张义正. 低密度脂蛋白受体/报告基因系统建立及中药降脂药物的筛选[J]. 天然产物研究与开发, 2005,17(3): 316
[5]张能锋, 祝宝华. 大鼠SERCA2a 启动子虫荧光素酶质粒的构建、鉴定和转染[J]. 实用临床医药杂志, 2006,10(2): 40
[6]Zamorano R, Suchindran S, Gainer JV. 3'Untranslated region of the type 2 bradykinin receptor is a potent regulator of gene expression [J]. Am J Physiol Renal Physiol,2006, 290(2):F456
[7]Li W,Gao L,Wang Y. Enhancement of cortisolinduced 11βhydroxysteroid dehydrogenase type 1 expression by interleukin 1β in cultured human chorionic trophoblast cells[J].Endocrinology,2006,147(5):2 490
[8]Ogawa R, Kagiya G, Kodaki T, et al. Construction of strong mammalian promoters by random cisacting element elongation[J].Biotechniques,2007,42(5):628
[9]Tsai CT, Wang DL, Chen WP, et al. Angiotensin II increases expression of alpha1C subunit of Ltype calcium channel through a reactive oxygen species and cAMP response elementbinding proteindependent pathway in HL1 myocytes[J]. Circ Res, 2007,100(10):1 476

备注/Memo

备注/Memo:
*河南省医学科技攻关基金资助项目2008002002#通讯作者,男,1965年生,教授,研究方向:肿瘤病因学,Email:zhaogq@zzu.edu.cn
更新日期/Last Update: 2010-05-28