Cloning and identification of fascin gene promoter region in esophageal cancer cells（克隆和鉴定fascin食道癌细胞基因启动子区域）.doc

Cloning and identification of fascin gene promoter region in esophageal cancer cells Author: Yang Zhangmin, Guo Zhangyan Gelatinase book Ying En-Min Li, Xie Jianjun, [Abstract] Objective: To clone identification fascin gene promoter region in esophageal cancer cells. Methods: application of PCR technology from esophageal cancer cell line EC109 amplified fragments of the the fascin gene 5 ‘flanking region of the 5 segments of different length, and firefly luciferase reporter gene was cloned into the expression vector pGL3 Basic (pGLB), then these plasmids and the control plasmid pGLB within the reference plasmid pRL TK co-transfected EC109 cells 48 h after the dual luciferase assay system analysis reorganization reporter gene transfected cells relative fluorescence intensity and combined bioinformatics analysis, to determine the location of the fascin gene promoter region. Results: The use of recombinant DNA technology successfully constructed five series of deletion recombinant luciferase reporter gene expression vector. The relative luciferase activity testing showed that compared with the control group, the above series of recombinant plasmid transfected cells showed a higher luciferase activity. bioinformatic analysis found that in the promoter region (-436 to +1) iconic regulatory elements TATA box (-34 to -29). Conclusions: fascin promoter region of the gene in esophageal cancer cells may be located in a section of about 436 bp upstream of the transcription start site. [Keywords] fascin the genes esophageal tumor promoter, Luciferases, reporter gene [Abstract] AIM: To clone and identify the promoter region of fascin gene in esophageal carcinoma cells. METHODS: Five dif ferent length fragments of 5 ‘flanking region of fascin gene were amplified from the genomic DNA of EC109 (an esophageal cancer cell line) by using PCR and cloned into luciferase reporter gene vector pGL3 Basic (pGLB, which aids in verification of functional promoter elements. Then