Yeast Sequencing Report
The ICL1 gene of Pichia pastoris, transcriptional regulation and use of its promoter
Article first published online: 4 SEP 2003
Copyright © 2003 John Wiley & Sons, Ltd.
Volume 20, Issue 13, pages 1097–1108, 15 October 2003
How to Cite
Menendez, J., Valdes, I. and Cabrera, N. (2003), The ICL1 gene of Pichia pastoris, transcriptional regulation and use of its promoter. Yeast, 20: 1097–1108. doi: 10.1002/yea.1028
- Issue published online: 4 SEP 2003
- Article first published online: 4 SEP 2003
- Manuscript Accepted: 22 JUN 2003
- Manuscript Received: 18 APR 2003
- Center for Genetic Engineering and Biotechnology, Cuba. Grant Number: 3082-327
- yeast Pichia pastoris;
- catabolite repression;
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 PICL1 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.