Project description:The goal of this set of experiments was to identify transcripts that are differentially expressed upon reactivation of NMD in an nmd2::HIS3 strain by galactose-induced expression of the NMD2 gene. Experiment Overall Design: Yeast strains harboring nmd2::HIS3 and the GAL1-NMD2 allele on a plasmid or the empty vector were grown in SC-medium containing raffinose but lacking uracile. Genome-wide expression profiles of the GAL-NMD2 and nmd2 deletion strains were analyzed over a 60-minute time course after adding galactose to the cell cultures. Four independent replicates were carried out for this experiment.
Project description:In response to carbon source switching from glucose to non-glucose, such as ethanol and galactose, yeast cells can directionally preprogram cellular metabolism to efficiently utilize the nutrients. However, the understanding of cellular responsive network to utilize a non-natural carbon source, such as xylose, is limited due to the incomplete knowledge on the xylose response mechanisms. Here, through optimization of the xylose assimilation pathway together with combinational evaluation of reported targets, we generated a series of mutants with varied growth ability. However, understanding how cells respond to xylose and remodel cellular metabolic network is far insufficient based on current information. Therefore, genome-scale transcriptional analysis was performed to unravel the cellular reprograming mechanisms underlying the improved growth phenotype.
Project description:Extensive transcriptional heterogeneity revealed by isoform profiling Application of TIF-Seq (Transcript IsoForm Sequencing) to S.cerevisiae. The method was applied to simultaneously identify the 5' capped mRNA transcription start site and the 3' polyadenylation site in different conditions: WT cells grown in glucose media [ypd, 2 biological replicates (bio) and 3 independent library preparations, technical replicates(lib)], WT cells grown in galactose media [ypgal, 4 biological replicates (bio) and 3 independent library preparations, technical replicates(lib)]. A modified protocol designed to enrich in long mRNA molecules was performed for WT cells grown in glucose media [ypd, 2 biological replicates (bio)] and in galactose media [ypgal, 2 biological replicates (bio)] conditions. Finally, control samples performed with a modified protocol designed to identify non-capped but polyadenylated molecules was performed in WT cells grown both in glucose (nypd) and galactose (nypgal) media.
Project description:We analyed the nucleosome positions by using 2 concentrations of micrococcal nuclease of yeast strains that were grown in raffinose and galactose containing media (synthetic complete).
Project description:Wild-type and isogenic H3K37R yeast cultures were grown in medium containing 2% raffinose as carbone source. Cells were synchronized in G1 with alpha factor. To half culture (100ml), glucose (2% final concentration) and 1.3ml BrdU (50mg/ml stock) and 1.3ml BrdU (50mg/ml stock) was added. To the other half, galactose (2% final concentration) and 1.3ml BrdU (50mg/ml stock). Both flasks were incubated for further 30min at 30C, then cell were collected by centrifugation and resuspended into 100ml of 30C warmed YPA-2%Glucose or YPA-2%Galactose medium containing 25ml HU (2M stock) plus 1.2 ml BrdU (50mg/ml stock) and incubated for 1 hour and 10minutes. YPA-Glucose and YPA-Galactose cultures were immediately transferred to ice/water bath and replication was stopped by addition of NaN3 and cells were processed for DNA immunoprecipitation with anti BrdU antibody.
Project description:This study was aimed to further illustrate the expression files of REN between glucose and raffinose in MRS broth. Transcriptomic analysis combined with mutants of the key genes based on homologous recombination technology indicated that galA1 gene cluster plays an important role in raffinose metabolism. Gene rafP and galA1 are responsible for raffinose transport and α-galactoside hydrolysis, followed by galactose hydrolysis by galKTE and sucrose hydrolysis by scrB. Lactobacillus salivarius Ren expanded the carbon utilization spectrum to adapt the fluctuating carbohydrate sources in the environment and shifted its carbohydrate metabolism to mixed-acid fermentation and then generated extra energy to bacterial growth when exposed to raffinose.