Jendraschak E, Kaminski W E
Department of Pathology, University of Washington School of Medicine, Seattle, Washington 98195, USA.
Genomics. 1998 May 15;50(1):53-60. doi: 10.1006/geno.1998.5290.
Knowledge of the promoter structure is critical for an understanding of the regulation of genes. We demonstrate by analysis of 405 human genes that human promoter regions are flanked by upstream Alu repeat elements, typically at a distance of 0.5-5 kb from their protein-coding areas. We identified common Alu repeat consensus sequences (ARC) among the different members of the Alu subfamilies that can be used as universal anchor sites for polymerase chain reaction (PCR) amplification. Utilizing ARC-specific primers and oligonucleotides specific for the 5' end of a selected target gene, we show that sequences spanning unknown human gene promoter regions can be directly amplified by PCR from genomic DNA. This novel technique, termed ARC-PCR, allowed us to characterize the proximal promoters of the human LTA4 hydrolase and SPARC genes, each within 1 day.
对启动子结构的了解对于理解基因调控至关重要。我们通过对405个人类基因的分析表明，人类启动子区域两侧是上游Alu重复元件，通常距离其蛋白质编码区域0.5-5 kb。我们在Alu亚家族的不同成员中鉴定出常见的Alu重复共有序列（ARC），可作为聚合酶链反应（PCR）扩增的通用锚定位点。利用针对选定靶基因5'端的ARC特异性引物和寡核苷酸，我们表明跨越未知人类基因启动子区域的序列可以通过PCR从基因组DNA中直接扩增。这种新技术称为ARC-PCR，使我们能够在1天内分别对人类LTA4水解酶和SPARC基因的近端启动子进行表征。
