Genomic structure analysis of SNC6, a progesterone-receptor associated protein gene, and cloning and characterization of its 5＇-flanking region
Genomic structure analysis of SNC6, a progesterone-receptor associated protein gene, and cloning and characterization of its 5'-flanking region作者机构：CancerInstituteSecondAffdiatedHospitalCollgeofMedicineZhefiangUniversityHangzhou310009China
出 版 物：《Journal of Zhejiang University Science》 (浙江大学学报（自然科学英文版）)
年 卷 期：2002年第3卷第1期
页 面：100-105页
核心收录：
学科分类：0710[理学-生物学] 07[理学] 071007[理学-遗传学]
基 金：863 High-Technnlo~ Project of China, (102-10-01-07) National Natural Science Foundation of China, NSFC, (39570785, 39770818)
主 题：SNC6 gene genomic structure 5'\|flanking region promoter luciferase assay
摘 要：Objective: To analyze the genomic structure of SNC6, a progesterone\|receptor associated protein gene and its regulatory elements in its 5 \|flanking region. Methods: Genomic sequence from GenBank database (accession number: Z98048) covering the whole SNC6 gene was used to analyze the genomic structure of SNC6 and design primers for PCR amplification of its 5 \|flanking region. A 1894 bp fragment of the 5 \|flanking region \{(-1814\} to +75) was cloned by PCR using genomic DNA from a healthy donor peripheral blood lymphocyte as template. This fragment, as well as 3 shorter derivative fragments (1423 bp, 632 bp and 416 bp, which correspond to -1344 to +75, -552 to +75 and -337 to +75 respectively), were subcloned into pGL2 series luciferase reporter vectors. These constructs were introduced into colorectal cancer cell line SW620 for transient expression of reporter gene and luciferase activities were measured. Results: The genomic structure analysis showed there are 12 exons for SNC6 gene, which spans 32017 bp (nt71529 to nt39513 in Z98048 sequence). All transfected SW620 cells with the above 5\|flanking region\|containing constructs showed luciferase activities. The highest luciferase activities were measured in transfected cells with vectors containing 1894 bp fragments, and the lowest luciferase activities were measured in transfected cells with vectors containing 416 bp fragments. Luciferase activities were higher in transfected cells with vectors containing 632 bp fragments than that in transfected cells with vectors containing 1423 bp fragments. Conclusion: The basic transcription\|promoting element (promoter) for SNC6 expression resides between 0 to -337, and two transcription\|enhancing elements (enhancer) resides between -337 to -552 and -1344 to -1814, whereas one transcription\|inhibiting element (silencer) exists between -552 to -1344.
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