The Preliminary Analysis And Appraisal Of STGC3gene Promoter

Objective: Using the bioinformatics and report gene vector analysis methods toclone and detect the activity of STGC3gene promoter region, preliminary identifiedthe promoter region of STGC3gene, provided the important evidence to clarify theregulation mechanism of STGC3gene expression.Methods: According to the preliminary findings of the STGC3gene, using severalbioinformatics software to conduct a length of3000bp promoter prediction, whichwas located in-3046bp to-46bp of STGC3gene5’upstream regulatory region.Constructed the possible promoter fragment to the vector of firefly luciferase reportergene pGL3-enhance, and then transfected the recombinant plasmid into humanembryonic kidney epithelial cell line293T, human nasopharyngeal carcinoma cellline CNE2and immortalized nasopharyngeal epithelial cell line NP69. In order topreliminary identify the gene promoter region of STGC3; we detected the activity ofthe recombinant plasmid by dual luciferase assay kit.Results: The results of bioinformatics analysis displayed that there were twotranscription start sites in the region, one was in-2348bp, and the other was in-948bp.This area had two CpG islands（CpG1and CpG2）, CpG1island，a length of101bp，was located in-1805bp to-1705bp; CpG2island，a length of217bp, waslocated in the-900bp to-674bp, which was not detected the classical array of TATAbox，but there was GC box between-1140bp and-774bp, and CAAT box in the-441bp.The promoter region of STGC3gene is located in－2992bp to－69bp.Thehighest score of promoter could reach the1.0in he region from-845bp to-795bp,thus，we speculate that this region may be the core area of the STGC3gene promoter.The experimental results showed that the pGL3-en283, pGL3-en281,pGL3-en571plasmid, especially the PGL3-en281promoter, had stronger promoter activity compared with the negative control pGL3-enhance plasmid in NP69、293Tand CNE2cells, and-934bp to-653bp may be the core promoter region.what is more,there may be some negative regulatory elements between-653bp and+72bp DNAsequence.The promoter activity of plasmid is stronger in293T than they were inCNE2cells, there may be some factors in CNE2cells.Conclusion:（1）STGC3gene basis of the promoter region is located in－2992bp to－69bp,and-934bp to-653bp may be the core promoter region.（2）There may be a negative regulatory element in the-653bp to+72bp DNAsequence of STGC3gene, and there were possible some factors thataffected the promoter activity in CNE2cells.