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Cloning and identification of fascin gene promoter region in esophageal carcinoma cells
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 19
- DOI:
- 10.3321/j.issn:1000-2790.2007.19.003
- Key Word:
- fascin基因;食管肿瘤;启动子;荧光素酶类;报告基因
Abstract： 目的:对食管癌细胞中fascin基因启动子区进行克隆鉴定. 方法:应用PCR技术从食管癌细胞系EC109中扩增出fascin基因5′侧翼区5段不同长度的片段,并将其克隆至萤火虫荧光素酶报告基因表达载体pGL3-Basic(pGLB)中, 然后将这些质粒与对照质粒pGLB、内参照质粒pRL-TK共转染EC109细胞,48 h后用双荧光素酶检测系统分析这些重组子报告基因转染细胞的相对荧光强度,并结合生物信息学分析,以判断fascin基因启动子区所在位置. 结果:用DNA重组技术成功构建了5个系列缺失重组荧光素酶报告基因表达载体. 相对荧光素酶活性检测表明,与对照组相比,上述一系列重组质粒转染细胞均显示出较高的荧光素酶活性. 生物信息学分析发现,在启动子区(-436～+1)有标志性调控元件TATA盒(-34～-29). 结论:在食管癌细胞中fascin基因的启动子区可能位于转录起始位点上游约436 bp的区段内.
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