An efficient method for isolating putative promoters and 5'-transcribed sequences from large genomic clones.

Abstract

Efficient strategies to isolate promoters and flanking exons from large genomic clones would facilitate the assembly of transcription units, complement existing techniques to isolate expressed sequences, and provide 5' regulatory elements. We have developed a rapid and simple method to isolate promoters from large mammalian genomic DNA clones by exploiting the abundance of binding sites for the ubiquitous transcription factor Sp1 in gene promoters. Using this method, putative promoter sequences with Sp1-binding sites are enriched approximately 100-fold from fragmented P1 clone DNA. Based on the abundance of Sp1-binding motifs in promoters, we predict that a significant subset of vertebrate promoters could be isolated by this method.