Project description:We report here the application of magnetic cell separation in microarray experiment to understand Drosophila gliogenesis during early embryogenesis. At stage 11, neuroectodermal cells were labeled by a fusion transmembrane protein, mCD8-GFP, in the wild-type embryos as well as in the embryos where glide/gcm was overexpressed. Following the cell dissociation, neuroectodermal cells were purified by using the commercial available anti-mCD8 microbeads. Microarray experiments based on prepared neuroectodermal cells from wild-type embryos verse glide/gcm overexpressed embryos at stage 11 have revealed 76 gene differentially regulated by overexpression of glide/gcm.
Project description:Drosophila melanogaster is a well-studied genetic model organism with several large-scale transcriptome resources. Here we investigate 7,952 proteins during the fly life cycle from embryo to adult and also provide a high-resolution temporal time course proteome of 5,458 proteins during embryogenesis. We use our large scale data set to compare transcript/protein expression, uncovering examples of extreme differences between mRNA and protein abundance. In the embryogenesis proteome, the time delay in protein synthesis after transcript expression was determined. For some proteins, including the transcription factor lola, we monitor isoform specific expression levels during early fly development. Furthermore, we obtained firm evidence of 268 small proteins, which are hard to predict by bioinformatics. We observe peptides originating from non-coding regions of the genome and identified Cyp9f3psi as a protein-coding gene. As a powerful resource to the community, we additionally created an interactive web interface (http://www.butterlab.org) advancing the access to our data.
Project description:Drosophila melanogaster is a well-studied genetic model organism with several large-scale transcriptome resources. Here we investigate 7,952 proteins during the fly life cycle from embryo to adult and also provide a high-resolution temporal time course proteome of 5,458 proteins during embryogenesis. We use our large scale data set to compare transcript/protein expression, uncovering examples of extreme differences between mRNA and protein abundance. In the embryogenesis proteome, the time delay in protein synthesis after transcript expression was determined. For some proteins, including the transcription factor lola, we monitor isoform specific expression levels during early fly development. Furthermore, we obtained firm evidence of 268 small proteins, which are hard to predict by bioinformatics. We observe peptides originating from non-coding regions of the genome and identified Cyp9f3psi as a protein-coding gene. As a powerful resource to the community, we additionally created an interactive web interface (http://www.butterlab.org) advancing the access to our data.
Project description:Chromatin accessibility mapping by DNase-seq on FACS-isolated cell populations during Drosophila melanogaster embryogenesis (6-8 hrs after egg-laying)
Project description:We report here the application of magnetic cell separation in microarray experiment to understand Drosophila gliogenesis during early embryogenesis. At stage 11, neuroectodermal cells were labeled by a fusion transmembrane protein, mCD8-GFP, in the wild-type embryos as wells as in the embryos where glide/gcm was overexpressed. Following the cell dissociation, neuroectodermal cells were purified by using the commercial available anti-mCD8 microbeads. Microarray experiments based on prepared neuroectodermal cells from wild-type embryos verse glide/gcm overexpressed embryos at stage 11 have revealed 76 gene differentially regulated by overexpression of glide/gcm. Among them, there are 11 genes have already been shown to be regulated by glide/gcm in the previous publications. Additionally, we have identified 4 more new glide/gcm-regulated genes by in situ hybridization or immunohistochemical staining. It is very convincing that reduction of the tissue complexity by cell purification leads to low false positives in microarray experiment. Keywords: other