Project description:In a previous study we adopted an integrated transcriptomic and proteomic approach to determine the physiological response of E. coli O157:H7 Sakai during exponential phase growth under steady-state conditions relevant to low temperature and water activity conditions experienced during meat carcass chilling in cold air (Kocharunchitt et al., 2012). The findings of that study provide a baseline of knowledge of the physiology of this pathogen, with the response of E. coli O157:H7 to steady-state conditions of combined cold and osmotic stress. To provide an insight into the genetic systems enabling this organism to adapt to growth at low temperature, we extended the aforementioned study to investigate the growth kinetics of E. coli O157:H7 Sakai during abrupt temperature downshift from 35 degrees C to 14 degrees C and, examined time-dependent global alterations in its genome expression upon cold shock from 35 degrees C to 14 degrees C. The genome-wide expression response of E. coli was analysed by both cDNA microarray (transcriptome response) and 2D-LC/MS/MS analysis (proteome response). Differences in gene and protein expression patterns in E. coli before and after cold shock were analysed through quantitative and comparative analysis of time series changes in both mRNA and proteins levels.
Project description:Comparative genome hybridization of transconjugants of E. faecalis OG1RF mated with V583. The total DNA of transconjugants was compared with wildtype strains to ascertain the amount of DNA that was transferred from E. faecalis V583 to E. faecalis OG1RF.
Project description:To isolate neuronal progenitor cells (NPCs), forebrains of E13.5 Miz1+/+ or Miz1-delta-POZ embryos were cut in small pieces, digested with trypsin and filtered through sterile gauze. Cells were cultivated in 2:1 DMEM/F12 supplemented with 1xB27 (Life technologies), 20 ng/ul EGF (Biomol), 20 ng/?l basic FGF (Biomol), 1 ug/ml fungizone (Gibco) and Penicillin/Streptomycin (PAA). NPCs were passaged every seven days. RNA expression of different genotypes was compared in sec. and quart. neurospheres.
Project description:Examine the dose response characteristics of a set of eight artificial transcripts (IVT) when spiked into a Universal Human RNA background at different copy numbers. The experiment was designed to <br>(i) evaluate the technical reproducibility of array data generated on the Agilent Whole Human 44K array with use of a set of external RNA controls <br> (ii) employ two panels of the same external RNA controls, which differed in copy number between panels, in order to simulate normal and disease states. We employed 8 different pools which were comprised of 8 different external RNA standards in a total human reference RNA background. Each different standard was present at a different copy number within any single pool (see file: E-MEXP-2670.additional.zip). Each control was present at a different concentration in each of the 8 pools. Standard microarray analysis was performed on the data and the differences in abundance between external RNA controls determined in terms of fold change differences.