Pheromone-inducible expression vectors for fission yeast Schizosaccharomyces pombe

Abstract

The fission yeast Schizosaccharomyces pombe is an attractive host for heterologous gene expression. However, expression systems for industrially viable large-scale fermentations are scarce. Several inducible expression vectors for S. pombe have been reported, with the strong thiamine-repressible nmt1⁺ promoter or derivatives thereof most commonly employed. Previously, the promoter regions of the genes sxa2⁺ and rep1⁺ were utilized to couple pheromone signaling to the expression of reporter genes for quantitative assessment of the cellular response to mating pheromones. Here, we exploit these promoters to serve as highly effective, plasmid-based inducible expression systems for S. pombe. Simply by adding synthetic P-factor pheromone, both promoters conferred 50-60% higher peak expression levels than the nmt1⁺ promoter. Full induction was significantly faster than observed for nmt1⁺-based expression platforms. Furthermore, the sxa2⁺ promoter showed very low basal activity and an overall 584-fold induction by synthetic P-factor pheromone. The dose-response curves of both promoters were assessed, providing the opportunity for facile tuning of the expression level by modulating P-factor concentration. Since the expression plasmids relying on the sxa2⁺ and rep1⁺ promoters require neither medium exchange nor glucose/thiamine starvation, they proved to be very convenient in handling. Hence, these expression vectors will improve the palette of valuable genetic tools for S. pombe, applicable to both basic research and biotechnology.

Keywords: Inducible expression system; P-factor; Pheromone response; Schizosaccharomyces pombe; rep1(+); sxa2(+).