Project description:NIH3T3 cells were cultured in medium containing 1 uM, 100 nM, 10 nM and 0 nM (serve as control) shld1 for 24 hours. Three independent cultures were obtained for each drug concentration. Total RNA from experimental samples were extracted using RNeasy Kit (Qiagen). Mouse RNA pooled from 11 cell lines (Stratagene, La Jolla, CA) was used as reference. Cy5-dUTP and Cy3-dUTP, were used to label the experimental cDNA and the reference samples, respectively, and hybridized to the Mouse Exonic Evidence Based Oligonucleotide (MEEBO) arrays (http://www.microarray.org./sfgf/meebo). The hybridizations were performed by Stanford Functional Genome Facility using its standard protocol (Oligo Array Hybridization protocol, http://www.microarray.org./sfgf/docView.do?type=2). A compound treatment design type is where the response to administration of a compound or chemical (including biological compounds such as hormones) is assayed. Computed
Project description:Caldicellulosiruptor saccharolyticus is an extremely thermophilic, Gram-positive anaerobe, which ferments cellulose-, hemicellulose- and pectin-containing biomass to acetate, CO2 and hydrogen. Its broad substrate range, high hydrogen-producing capacity, and ability to co-utilize glucose and xylose, make this bacterium an attractive candidate for microbial bioenergy production. Glycolytic pathways and an ABC-type sugar transporter were significantly up-regulated during growth on glucose and xylose, indicating that C. saccharolyticus co-ferments these sugars unimpeded by glucose-based catabolite repression. The capacity to simultaneously process and utilize a range of carbohydrates associated with biomass feedstocks represents a highly desirable feature of a lignocellulose-utilizing, biofuel-producing bacterium. Keywords: substrate response C. saccharolyticus was subcultured (overnight) 3 times on the substrate of interest in modified DSMZ 640 medium before inoculating a pH-controlled (pH = 7) 1-liter fermentor containing 4 gram substrate per liter. Cells were grown at 70 °C until mid-logarithmic phase (~OD660 = 0.3-0.4) and harvested by centrifugation and rapid cooling to 4 °C and stored at -80 °C. To elucidate the central carbon metabolic pathways and their regulation, transcriptome analysis was performed after growth on glucose, xylose and a 1:1 mixture of both substrates. L-Rhamnose, which was likely to follow another pathway, was used as a reference substrate.
Project description:The aim of this study was to conduct a genome-wide analysis for constituent tuber carotenoid QTL. Using a genetical genomics approach samples from clones with similar carotenoid traits were bulked and patterns of gene expression were measured for each bulk by microarray analysis. Variation of gene expression within these bulks may be due to either polymorphisms located near to or within the gene (cis-eQTL) or indirectly from a distant location on the genome (trans-eQTL). Differentially expressed clones from bulks with contrasting carotenoid traits were genetically mapped in order to re-enforce the QTL analysis, and provide a rapid means of developing gene markers closely associated with the target traits.
Project description:Gene expression profiling in leaves of a free-growing aspen tree (Populus tremula) in Umea in northern Sweden during natural autumn senescence (from August 17 to September 21).
Project description:Analysis of transcriptional response to UV irradiation in two related crenarchaea, Sulfolobus solfataricus and Sulfolobus acidocaldarius.
Project description:Exponentially growing Sulfolobus acidocaldarius were treated with NaAc to generate replication runout and arrest in G2 phase. The cells were then resuspended in fresh acetate-free media which generates a synchronous population. Samples for investigation of gene expression change were taken during the synchronised populations progress through the cell cycle.
Project description:Transcript-Level Variation in Barley Seedling Leaves Challenged with Puccinia hordei and the Molecular Basis of Partial Resistance to Leaf Rust
Project description:Transcript-Level Variation in Barley Seedling Leaves Challenged with Puccinia hordei and the Molecular Basis of Partial Resistance to Leaf Rust
Project description:Regulation of gene expression underlies the establishment and maintenance of cell identity. Chromatin structure and gene activity are linked. Recently CTCF anchored loops have been described as major features of chromatin organisation. However, the dynamics and role for these structures in differentiation is unknown. We used Tethered Chromatin Conformation Capture (TCC) to assess for the dynamics of CTCF-anchor loop formation upon differentiation of mouse embryonic stem cells (ESC) and neural stem cells (NSC).
Project description:We have compared gene expression in human nasal brushing cells from 19 cystic fibrosis (CF) patients and 19 healthy controls using a 5.2K cDNA microarray. Our aim is to identify new disease biomarkers for the Cystic Fibrosis Gene Therapy Consortium. These markers will be used to report more effectively on the response to the administration of gene therapy in vivo. Cystic Fibrosis is a recessive genetic disease caused by mutations in the cystic fibrosis conductance regulator (CFTR) gene which encodes a chloride ion channel. The most common mutation is the ∆F508 mutation, present on 70% of CF chromosomes in Caucasian populations. The disease affects many organs in the body such as the pancreas, liver, sweat glands, small intestine and reproductive tracts but is most commonly associated with progressive, inflammatory lung disease. The current average life expectancy of CF patients is 35 years. Gene therapy is being developed as a treatment for CF airway disease, however, means of measuring the efficiency and efficacy of gene therapy in vivo are lacking. This is mainly due to the difficulty in measuring the chloride conductance of CFTR in cells and tissues. Furthermore, clinical assays for measuring improvements in lung function are insensitive. Surrogate markers of inflammation and CFTR function will therefore be important for the effective assessment of gene therapy in vivo. We have analysed gene expression in human nasal epithelium as this is considered an accessible surrogate for the conducting airways where disease manifests in the majority of patients. Additionally, this tissue will be sampled in clinical trials.