Ura− host strains for genetic manipulation and heterologous expression of Torulaspora delbrueckii

Recently, the industrial and academic interest in the yeast Torulaspora delbrueckii has increased notably due to its high resistance to several stresses. This characteristic has made of this organism a very attractive model to study the molecular basis of the stress response in yeast. However, very little is known about the physiology and genetics of this yeast, and the tools for its manipulation have not been developed. Here, we have generated Ura− strains of the bakerʹs yeast T. delbrueckii IGC5323 by either 5-FOA-aided selection or transformation with a PCR-based disruption cassette, natMX4, which confers nourseothricin resistance. Furthermore, the mutant and disruptant strains were used as recipient of a plasmid containing the xlnB cDNA from Aspergillus nidulans. Our results indicate that Torulaspora transformants produce active recombinant protein at a similar level to that found for Saccharomyces.