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Molecular cloning of a cDNA encoding enolase from the filamentous fungus, Aspergillus oryzae
M Machida1, Y C Chang, M Manabe
1Department of Molecular Biology, National Institute of Bioscience and Human-Technology, Higashi 1-1, Tsukuba, Ibaraki 305, Japan.
Abstract:
A 1.6-kbp full-length cDNA for the Aspergillus oryzae enolase gene (enoA) was cloned. The sequenced insert contained a continuous open reading frame of 1314 bp encoding a protein of molecular weight 47 405. Among all enolases sequenced to-date, the deduced amino-acid sequence showed the highest homology (74.9%) with Candida albicans enolase (ENO1). Strong codon biases and multiple transcription start sites downstream from CT-blocks in the 5'-flanking region suggested strong expression. enoA mRNA was found to occupy approximately 3% (w/w) of total mRNA of A. oryzae by quantitative RT-PCR. This strong transcription was dependent on the carbon source in the medium and correlated with the growth rate of the mycelium.