Project description:We developed an artificial genome evolution system, which we termed ‘TAQing’, by introducing multiple genomic DNA double-strand breaks using a heat-activatable endonuclease in mitotic yeast. The heat-activated endonuclease, TaqI, induced random DSBs, which resulted in diverse types of chromosomal rearrangements including translocations. Array comparative genomic hybridization (aCGH) analysis was performed with cell-fused Saccharomyces cerevisiae strains induced genome evolution by TAQing system. Some of copy number variations (CNVs) induced by massive genome rearrangements were detected in the TAQed yeast strains.
Project description:Model-guided chassis strain design has the potential to accelerate cellfactory development. In this experiment genetic targets were identified in silico and implemented in vivo to design a yeast chassis strain for enhanced production of succinic, malic and fumaric acid. The phenotype of engineered chassis strains was further optimised through adaptive laboratory evolution. RNA-seq analysis of engineered yeast chassis strains, evolved strains and wild-type (CEN.PK background)was performed to determine the effect of engineered gene deletions and evolution on the transcriptome.
Project description:We constructed S. cerevisiae BY_DEH+ strain which is able to assimilate both 4-deoxy-L-erythro-5-hexoseulose uronate (DEH, a monouronic acid produced by digestion of alginate with exo-type alginate lyase) and mannitol from BY4742 strain and improved its ability to assimilate DEH through an adaptive evolution (Matsuoka et al. Sci. Rep. 2017, 7, 4206). To examine transcriptional responses of the yeast to DEH and mannitol, gene expressions of the evolved strain (BY_DEH++ strain) in DEH medium, mannitol medium, and glucose medum were analyzed. For revealing the mechanisms underlying the adaptive evolution, gene expressions of both BY_DEH+ strain and BY_DEH++ strain in both DEH medium and glucose medium were measured.
