[1]杨聚荣,何娅妮,张剑凯,等.人NaDC1基因启动子序列转录调控元件的初步分析[J].第三军医大学学报,2007,29(18):1787-1789.
 YANG Ju-rong,HE Ya-ni,ZHANG Jian-kai,et al.Positional identification of transcription regulation elements in human NaDC1 promoter[J].J Third Mil Med Univ,2007,29(18):1787-1789.
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人NaDC1基因启动子序列转录调控元件的初步分析(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第18期
页码:
1787-1789
栏目:
论著
出版日期:
2007-09-30

文章信息/Info

Title:
Positional identification of transcription regulation elements in human NaDC1 promoter
作者:
杨聚荣何娅妮张剑凯李新伦梁海君林静吴小玮王惠明李开龙
第三军医大学大坪医院野战外科研究所肾内科;广东医学院基础学院人体解剖学教研室
Author(s):
YANG Ju-rong HE Ya-ni ZHANG Jian-kai LI Xin-lun LIANG Hai-jun LIN JingWU Xiao-wei WANG Hui-ming Li Kai-long
Department of Nephrology, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042;Department of Anatomy, College of Basic Medicine, Guangdong Medical College, Zhanjiang 524023, China
关键词:
hNaDC1基因双荧光素酶报告系统转录调控
Keywords:
human Na+ dependent dicarboxylate transporter 1  gene report gene regulation of gene expression
分类号:
R394-33;R394.2
文献标志码:
A
摘要:
目的 对hNaDC1基因启动子序列转录调控元件进行初步分析。 方法 构建hNaDC1基因5′侧翼序列系列缺失质粒,以双荧光素酶报告基因检测系统检测hNaDC1基因5′侧翼区(-2 232/+136)的转录调控元件与转录调控蛋白之间的相互作用。 结果 pGL3-hNaDC1A质粒的转录活性最高,为(2.98±0.45);pGL3hNaDC1D的转录活性最低,为(0.45±0.14)。以pGL3hNaDC1A质粒的转录活性为基准(100%),其他4种质粒相应的转录活性分别为:pGL3-hNaDC1B 86.7%,pGL3-hNaDC1C 89.4%,pGL3-hNaDC1D 15.8%,pGL3-hNaDC1E 61.2%。MatInspector 5.0软件预测结果显示hNaDC1基因5′侧翼启动子序列共含有22个14种转录因子结合位点。 结论 hNaDC1基因5′侧翼区-1 084/-254和-12/+136区域可能存在有正性作用转录因子的结合位点。
Abstract:
Objective    To determine the positions of transcription regulation elements in 5’ flanking region of human Na+ dependent dicarboxylate transporter 1 (hNaDC1) gene.     Methods    The DNA fragments hNaDC1 A-E(-2 232/+136)of its 5’ flanking were amplified from the genomic DNAs of HK-2 cells by using PCR. The PCR products were directionally subcloned into pGL3-basic vector. The recombined clones were identified by agarose gel electrophoresis after restriction endonuclease digesting and DNA sequencing. The recombined vectors as well as pGL3-basic vector were respectively transfected into HK-2 cells with Lipofectamine 2000. To normalize transfection efficiencies, 0.4 μg of a Renilla luciferase positive control vector (pRL-CMV plasmid) was used for co-transfection.The interaction between transcription regulation elements and transcription regulation proteins in 5’ flanking region of hNaDC1 gene(-2 232/+136)was measured by analyzing relative light unit (RLU) with the help of Dual Luciferase Report Gene Assay System (DLR).     Results    The RLU was significantly lowered in HK-2 cells transfected with pGl3-basic when compared with the cells harboring pGL3-hNaDC1 A-E (P<0.05 or P<0.001). Compared with pGL3-hNaDC1 A group, RLU was significantly lowered in pGL3-hNaDC1 D group (P<0.01) and pGL3-hNaDC1 E group (P<0.05). MatInspector 5.0 system indicated that there were 22 binding sites of 14 trans-acting factors(Sore>90) in the regions -1 084/-254 and -12/+136.     Conclusion    Transcription enhancer elements, which have interacted with transcription activation proteins, exist in both region -1 084/-254 and -12/+136. These results may offer an approach for further study on the enhancers of hNaDC1 gene.

相似文献/References:

[1]杨聚荣,张剑凯,何娅妮,等.人NaDC1基因近端启动子生物信息学分析及载体构建[J].第三军医大学学报,2007,29(14):1410.
 YANG Ju-rong,ZHANG Jian-kai,HE Ya-ni,et al.Bioinformatics analysis of human NaDC1 proximal promoter and construction of firefly report gene vector[J].J Third Mil Med Univ,2007,29(18):1410.

更新日期/Last Update: 2008-07-16