Project description:This experiment was performed to measure differences in pathway-level evolution of cis-regulation between closely related Kluyveromyces species. Sequencing of mRNA as well as DNA was performed in hybrids of Kl. lactis with two other species, Kl. wickerhamii and Kl. marxianus. Cells were grown overnight for DNA sequencing and for 4-8h to an OD600=0.7-1.0 for mRNA sequencing. DNA and mRNA reads were mapped to hybrid genomes, multimapping reads discarded, and allele-specific expression ratio for each gene was calculated after first normalizing mRNA reads to the number of DNA reads for each gene in each species. We found that the ribosomal protein genes showed pathway-level allele-specific expression differences between species in both interspecies hybrids.
Project description:Total RNA versus genomic DNA hybridization on custom arrays designed for all Kluyveromyces waltii genes Total RNA was collected in mid-log phase from Kluyveromyces waltii 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 waltii.
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.