Lin Gong-biao, Xiao Jian-yun, Qiu Yuan-zheng, Wang Cheng-long, Tian Yong-quan, Zhao Su-ping
Department of Otorhinolaryngology, the First Hospital, Fujian Medical University, Fuzhou, Fujian, PR China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2004 Feb;21(1):35-8.
To investigate the mechanism of cytokeratin 13 (CK13) gene expression control and the effects of different motifs of CK13 gene 5' flanking region on its transcriptional activity.
The molecular clone technique and reporter gene analysis were used to assay the effects of different motifs of 513 bp of CK13 gene 5' flanking region on its transcriptional activity. The pCAT enhancer vectors with different motifs of CK13 gene 5' flanking region were constructed and transferred to HeLa cells with the help of lipofectin. The instant CAT expression of different clones was detected and the effects of different motifs of the CK13 gene 5' flanking region on its transcriptional activity were evaluated.
119 bp from -nt.325 to -nt.207 upstream of the first ATG of CK13 gene 5' flanking region included a silent element. 113 bp region from -nt.206 to -nt.94 included an enhanced element.
513 bp of CK13 gene 5' flanking region includes a silent element and an enhanced element. Further locating these cis elements and detecting the related trans reaction factors may unveil some important clues to the details of the mechanisms for the CK13 gene expression and tissue-specific expression.
研究细胞角蛋白13(CK13)基因表达调控机制以及CK13基因5′侧翼区不同基序对其转录活性的影响。
采用分子克隆技术和报告基因分析法,检测CK13基因5′侧翼区513 bp不同基序对其转录活性的影响。构建含CK13基因5′侧翼区不同基序的pCAT增强子载体,借助脂质体转染HeLa细胞。检测不同克隆的瞬时CAT表达,评估CK13基因5′侧翼区不同基序对其转录活性的影响。
CK13基因5′侧翼区第一个ATG上游-nt.325至-nt.207的119 bp包含一个沉默元件。-nt.206至-nt.94的113 bp区域包含一个增强元件。
CK13基因5′侧翼区513 bp包含一个沉默元件和一个增强元件。进一步定位这些顺式元件并检测相关反式作用因子,可能为CK13基因表达及组织特异性表达机制的细节揭示一些重要线索。