Identification of a sequence containing the positive regulatory region of Saccharomyces cerevisiae gene ENO1.
A DNA fragment which contains the 5'-flanking region of the Saccharomyces cerevisiae enolase 1 gene (ENO1) and a portion of the coding sequence was cloned in a plasmid pMC1587. This fragment was fused in frame to the lacZ gene of Escherichia coli. Many mutants which deleted a portion of the 5'-flanking region of ENO1 were isolated from this ENO1-lacZ fusion plasmid by in vitro recombination. Analysis of beta-galactosidase activity of these mutants indicated that the regulatory region responsible for an efficient expression of the ENO1-lacZ fused gene resides within an 86-bp sequence located at -487 to -402 upstream from the start codon of ENO1. We found that the segment encompassing the 86-bp region worked equally well in an inverted orientation, but the tandem duplication of the sequence did not enhance the expression of the fused gene.
D001426:Bacterial Proteins / Q000096:biosynthesis; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004274:DNA, Recombinant; D005798:Genes, Bacterial; D005809:Genes, Regulator; D010751:Phosphopyruvate Hydratase / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D010751:Phosphopyruvate Hydratase