Cloning and Nucleotide Sequence of the Alkaline Protease Gene fromFusariumsp. S-19-5 and Expression inSacchavomyces cerevisiae

摘要

We have cloned a genomic DNA encoding the alkaline protease (Alp) ofFusariumsp. S-19-5 from a genomic DNA library and sequenced the nucleotides. Complementary DNA encoding Alp was also isolated from the cDNA library after amplifying the gene by PCR using partial sequences of the Alp genomic DNA as primers. The Alp gene has an open reading frame of 1137 nucleotides containing three introns. A TATA box (TAAATA) was observed 112 base pairs upstream from the translation initiation codon in the 5′-non coding region. The Alp protein has a pre region consisting of 14 amino acids and a pro region of 85 amino acids preceding the mature region, which consists of 280 amino acids. The amino acid sequence ofFusariumAlp has 52 homology with that ofAspergillus oryzaeand 51 homology with that ofAcremonium chrysogenum. The entire cDNA encodingFusariumAlp was introduced intoSaccharomyees cerevisiae, which then secreted enzymatically active Alp into the culture medium.