Project description:Understanding the genotype-phenotype map and how variation at different levels of biological organization is associated are central topics in modern biology. Fast developments in sequencing technologies and other molecular omic tools enable researchers to obtain detailed information on variation at DNA level and on intermediate endophenotypes, such as RNA, proteins and metabolites. This can facilitate our understanding of the link between genotypes and molecular and functional organismal phenotypes. Here, we use the Drosophila melanogaster Genetic Reference Panel and nuclear magnetic resonance (NMR) metabolomics to investigate the ability of the metabolome to predict organismal phenotypes. We performed NMR metabolomics on four replicate pools of male flies from each of 170 different isogenic lines. Our results show that metabolite profiles are variable among the investigated lines and that this variation is highly heritable. Second, we identify genes associated with metabolome variation. Third, using the metabolome gave better prediction accuracies than genomic information for four of five quantitative traits analyzed. Our comprehensive characterization of population-scale diversity of metabolomes and its genetic basis illustrates that metabolites have large potential as predictors of organismal phenotypes. This finding is of great importance, e.g., in human medicine, evolutionary biology and animal and plant breeding.
Project description:Absolute (molar) quantification determines proteins stoichiometry in complexes, networks and metabolic pathways. We employed MS Western workflow to determine molar abundances of proteins critical for morphogenesis and phototransduction (PT) in eyes of Drosophila melanogaster using a single chimeric 264 kDa protein standard that covers, in total, 197 peptides from 43 proteins. Each protein was independently quantified with 2 to 4 proteotypic peptides with the coefficient of variation of less than 15 %, better than 1000-fold dynamic range and sub-femtomole sensitivity. We determined molar abundances and stoichiometric ratios of the components of the PT machinery and the rhabdomere, and how they are changing when rhabdomere morphogenesis is perturbed by genetic manipulation of the evolutionary conserved gene crumbs (crb).
Project description:Drosophila melanogaster is a validated eukaryotic model for immunity-concerned studies in the post-genomic era. In the present study we performed oral experimental infection of D. melanogaster with Pseudomonas aeruginosa (strain ATCC27853). By using a whole genome microarray approach, we intended to identify significant alterations in the expression profile of relevant genes amenable to qualify as new models for the investigation of specific host-parasite interactions.