Cloning of the glyceraldehyde-3-phosphate dehydrogenase gene from Pseudozyma flocculosa and functionality of its promoter in two Pseudozyma species

Abstract

Pseudozyma flocculosa is a yeast-like epiphyte recently classified as a basidiomycete related to the Ustilaginales. In this study, we report the cloning of its gene coding for a putative glyceraldehyde-3-phosphate dehydrogenase (GPD). This gene was selected on the premise that its transcripts are abundant during the growth phase of P. flocculosa. The complete sequence of this gene was found to contain two introns in the coding region and one in the 3'-untranslated region. This gene was present in a single copy in the genome of P. flocculosa. By comparing its deduced amino acid sequence with various sequences from basidiomycetous and ascomycetous fungi, we observed a stronger homology with the former group as predicted by the new classification of P. flocculosa. The promoter region lacked a typical TATA or CAAT box but contained a CT-rich region including the transcription start site. Although the GPD promoter showed a stronger affinity within P. flocculosa, it remained active across species as shown by expressing the green fluorescent protein in Pseudozyma antarctica. The cloning of this gene and its promoter brings new and versatile options to the limited genetic tools currently available for the study of the recently defined Pseudozyma genus.