Premium
The ICL1 gene of Pichia pastoris , transcriptional regulation and use of its promoter
Author(s) -
Menendez Javier,
Valdes Iris,
Cabrera Nelson
Publication year - 2003
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1028
Subject(s) - pichia pastoris , isocitrate lyase , biology , microbiology and biotechnology , open reading frame , gene , saccharomyces cerevisiae , northern blot , pichia , genomic library , southern blot , yeast , peptide sequence , genetics , gene expression , biochemistry , recombinant dna , glyoxylate cycle , enzyme
We cloned and characterized a gene encoding isocitrate lyase from the methylotrophic yeast Pichia pastoris . This gene was isolated from a P. pastoris genomic library using a homologous PCR hybridization probe, amplified with two sets of degenerate primers designed from conserved regions in yeast isocitrate lyases. The cloned gene was sequenced and consists of an open reading frame of 1563 bp encoding a protein of 551 amino acids. The molecular mass of the protein is calculated to be 60.6 kDa with high sequence similarity to isocitrate lyase from other organisms. There is a 64% identity between amino acid sequences of P. pastoris Icl and Saccharomyces cerevisiae Icl. Northern blot analyses showed that, as in S. cerevisiae , the steady‐state ICL1 mRNA levels depend on the carbon source used for cell growth. Expression in P. pastoris of the dextranase gene ( dexA ) from Penicillium minioluteum under control of the ICL1 promoter proved that P ICL1 is a good alternative for the expression of heterologous proteins in this methylotrophic yeast. The sequence presented here has been deposited in the EMBL data library under Accession No. AJ272040. Copyright © 2003 John Wiley & Sons, Ltd.