摘要
目的:克隆端粒、端粒酶结合因子hPinx1基因的启动子,分析并鉴定其活性调控元件。方法:采用PCR技术从人肝癌细胞系HepG2基因组中扩增出hPinx1启动子,构建到萤光素酶报告基因载体pGL3-basic中,确定所扩增的DNA序列,在HepG2细胞中检测其活性。结果:克隆了hPinx1基因转录起始位点上游4661bp且序列正确;活性分析表明hPinx1启动子含有多个调控元件,其中核心序列位于530bp内,在1329～2174bp间存在正调控序列,在2174～4661bp间存在负调控序列。结论:构建的hPinx1启动子具有活性,为hPinx1的功能研究提供了重要基础。
Objective： To clone telomere/telomerase binding factor hPinxl promoter and insert it into a luciferase reporter gene vector, and to characterize the promoter activity. Methods： The promoter of hPinxl was amplified from human liver cancer cell line HepG2 genomic DNA by PCR and constructed into pGL3-basic vector. The recombinant plasmid was determined by DNA sequencing. The promoter activity was analyzed in HepG2 cell line. Results： The upstream 4 661 bp of hPinxl transcriptional start site was constructed successfully. The hPinxl promoter contained many regulatory elements. The core region of hPinxl promoter located witnin 530 bp, there were a positive regulatory sequence from 1 329 to 2 174 bp , and a negative regulatory sequence from 2 174 to 4 661 bp. Conclusion： The activity assay confirmed that the sequence had promoter activity. This is important for hPinxl further function investigation.
出处
《生物技术通讯》
CAS
2008年第5期645-648,共4页
Letters in Biotechnology
