Fig. 1
From: Engineering prokaryotic regulator IrrE to enhance stress tolerance in budding yeast
A general schematic for the directed evolution of IrrE to improve microbial tolerance. The strategy of error-prone PCR was carried out to introduce diversity to the genes. The mutant genes are then ligated into the pRS416 vector with HXT7 promoter and TEF1 terminator and transferred into S. cerevisiae BY4742. The library is screened with a high-throughput method based on the growth biomass in the presence of FAP. High biomass is related with faster growth and selected for further analysis in flask fermentation. Improved mutants are isolated for the next round of modification and screening. According to the transcriptome analysis, regulatory networks were constructed to reveal the regulatory mechanisms by which IrrE and I24 enhance yeast tolerance