“-GCT(A)GCGAGATC-”序列负向调控Nanog基因启动子-135/-230区活性研究  

Activity of Nanog promoter -135/-230 could be reverse-regulated by "-GCT( A) GCGAGATC-" sequence

作　　者：杜娟[1,2] 吴博 殷松娜[1] 王爱红 庞秋霞 赵菊梅[1,2] DU Juan;WU Bo;YIN Song-na;WANG Ai-hong;PANG Qiu-xia;ZHAO Ju-mei(Medical College of Yan＇an University,Yan＇an 716000,China;Yan＇an Key Laburatory of the Prevention and Treatment of Tumor,Yan＇an 716000,China)

机构地区：[1]延安大学医学院,陕西延安716000 [2]延安市肿瘤防治研究重点实验室,陕西延安716000

出　　处：《延安大学学报（医学科学版）》2018年第3期1-6,12,共7页Journal of Yan'an University:Medical Science Edition

基　　金：陕西省青年人才托举计划项目(20160223);延安市博士科研启动资金资助(205040125)

摘　　要：目的探究Nanog启动子-332/+50区与双荧光素酶报告载体p GL4. 10连接后负向调节Nanog-135/-230区活性的调节序列。方法截短法将Nanog启动子-812-+171区不同片段与p GL4. 10载体连接,脂质体转染法将各载体分别与内参载体p RL-TK共转入细胞,双荧光素酶系统报告系统检测报告基因的表达变化。结果Nanog启动子-812/-332区、+50/+171区及-332/+51(或+56)区均不含Nanog-135/-230区的负向调控元件,改变Naong-332/+50区与报告载体p GL4. 10的连接位点(NheⅠ,BglⅡ)行成序列"-GCT (A)GCGAGATC-"即可消除Nanog启动子-332/+50对报告基因的正向调控作用。结论待检测启动子与载体连接构成"-GCT(A) GCGAGATC-"序列后可负向调控Nanog-135/-230区的活性。Objective Identify the regulatory sequence of Nanog promoter -332/＋50 that down-regulates Nanog -135/-230＇s activity. Methods Construct a set of plasmids that ligate different Nanog promoter -812-/＋171 truncates with pGL4.10, co-transfect pRL-TK with the plasmid minded to cells using Lipofectamine 2000, testing the promoter activity using the Dual Luciferase Reporter assay system. Results There have no regulating sequence in the Nanog promoter -812/-332, ＋50/＋171, and -332/＋51. A change of ligated sequence between the NheⅠ, BglⅡ restriction sites maintained in pGL4.10 and Nanog promoter -332/＋50 sequence, “-GCT（A）GCGAGATC-”, could counteract the regulatory effect of Nanog promoter -332/＋50 on report gene expression. Conclusion Sequence of “-GCT（A）GCGAGATC-” could reversely regulate Nanog 332/＋50 promoter activity.

关 键 词：启动子报告载体构建 隐蔽元件 修饰元件 转录调节 

分 类 号：R34[医药卫生—基础医学]