Cloning and characterization of a novel NAD+‐dependent glyceraldehyde‐3‐phosphate dehydrogenase gene from Candida glycerinogenes and use of its promoter. Issue 4 (12th March 2013)

• Record Type: Journal Article
• Authors: Zhang, Cheng; Zhuge, Bin; Zhan, Xiaobei; Fang, Huiying; Zong, Hong; Zhuge, Jian
• Is Part Of: Yeast. Volume 30:Issue 4(2013:Apr.)
• Page Start: 157
• Page End: 163
• Publication Date: 2013-03-12
• Subjects: Yeast -- Periodicals; Yeasts -- Periodicals; Yeasts -- genetics -- Periodicals; Electronic journals; 547
• DOI: 10.1002/yea.2946
• Languages: English
• ISSNs: 0749-503X

Abstract

<abstract abstract-type="main"> <title>Abstract</title> <p>A 3950 bp genomic fragment from <italic>Candida glycerinogenes</italic>, WL2002‐5, containing the Cg<italic>GAP</italic> gene encoding a glyceraldehyde‐3‐phosphate dehydrogenase homologous to GAP genes in other yeasts using degenerate primers, was cloned and characterized with inverse PCR. Sequence analysis revealed a 1164 bp open reading frame encoding a putative peptide of 387 deduced amino acids, with a molecular mass of 36 kDa. The Cg<italic>GAP</italic> protein consisted of an N‐terminal NAD⁺‐binding domain and a central catalytic domain. Six stress‐response elements were found in the upstream region of the Cg<italic>GAP</italic> gene. The influence of Cg<italic>GAP</italic> on glycolysis was investigated. Functional analysis revealed that <italic>Saccharomyces cerevisiae</italic> transformed with Cg<italic>GAP</italic> was restored to the wild‐type phenotype when cultured in high‐osmolarity medium, suggesting that it is a functional <italic>GAP</italic> protein. Promoter studies in <italic>S</italic>. <italic>cerevisiae</italic> using the green fluorescent protein (<italic>gfp</italic>) gene as a reporter showed that the <italic>GAP</italic> promoter (P_{Cg<italic>GAP</italic>}) is constitutively expressed in <italic>S</italic>. <italic>cerevisiae</italic> cells grown on glucose. Copyright © 2013 John Wiley &amp; Sons, Ltd.</p> </abstract>