Project description:The purpose of this experiment was to identify the genes bound by Ndt80 in K. lactis when constitutively expressed. Ndt80 was tagged with c-myc, the native Ndt80 promoter was replaced with the K. lactis Gal1 promoter and the protein was immunoprecipitated with a c-myc antibody. Cells were grown overnight in SRaffinose until log-phase, then in SRaffinose + 1.7% Galactose for 5 hours. Each experiment was repeated twice and sequenced on an Illumina HiSeq 4000.
Project description:The purpose of this experiment was to identify the genes bound by Ndt80 in S. cerevisiae when constitutively expressed, for the native Ndt80 as well as a heterologous Ndt80 from Pichia pastoris. For the ChIPs on the native Ndt80s, Ndt80 was tagged with c-myc; for the heterologous ChIP P. pastoris Ndt80 was integrated into the S. cerevisiae genome and tagged with c-myc. In both cases, the native Ndt80 promoter was also replaced with an A. gossypii Gal1 promoter and the protein was immunoprecipitated with a c-myc antibody. Cells were grown overnight in SRaffinose until log-phase, then in SRaffinose + 1.7% Galactose for 5 hours. Each experiment was repeated twice and sequenced on an Illumina HiSeq 4000.
Project description:The purpose of this experiment was to identify the genes bound by Ndt80 in P. pastoris when constitutively expressed. Ndt80 was tagged with c-myc, the native Ndt80 promoter was replaced with the E. gossypii Tef1 promoter and the protein was immunoprecipitated with a c-myc antibody. Cells were grown unti log-phase in YPD. Each experiment was repeated twice and sequenced on an Illumina HiSeq 2500.
Project description:Total RNA versus genomic DNA hybridization on custom arrays designed for all Kluyveromyces lactis genes Total RNA was collected in mid-log phase from Kluyveromyces lactis cells grown in rich medium (abbreviated CM, in house recipe). RNA was then converted to cDNA, Cy3-labeled and hybridized competitively against Cy5 labeled genomic DNA from Kluyveromyces lactis.
