Isolation of human promoter regions by Alu repeat consensus-based polymerase chain reaction.
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.
Jendraschak
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Department of Pathology, University of Washington School of Medicine, Seattle, Washington 98195, USA.
Kaminski
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WE
D020087:Alu Elements / Q000235:genetics*; D001483:Base Sequence; D016384:Consensus Sequence / Q000235:genetics*; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D005784:Gene Amplification; D006801:Humans; D017572:Leukotriene A4 / Q000235:genetics; D008969:Molecular Sequence Data; D015676:Osteonectin / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA
D017572:Leukotriene A4; D015676:Osteonectin; D004247:DNA