Project description:Alphaviruses establish a persistent infection in arthropod vectors, which is essential for effective transmission of the virus to vertebrate hosts. The development of persistence in insects is not well understood, although it is thought to involve the innate immune response. Using a transgenic fly system (SINrep) expressing a self-replicating viral genome, we have previously demonstrated the antiviral response of the Drosophila Imd (Immune Deficiency) and Jak-STAT innate immunity pathways. In the current study, microarray analysis of SINrep flies in comparison to control GFP flies aims to detect genes that are sensitive to Sindbis viral RNA replication. Both SINrep and GFP adult flies were harvested 3 days post eclosion for RNA extraction and hybridization on Affymetrix microarrays. We look for genes significantly altered in the presence of viral RNA replication.
Project description:Alphaviruses establish a persistent infection in arthropod vectors, which is essential for effective transmission of the virus to vertebrate hosts. The development of persistence in insects is not well understood, although it is thought to involve the innate immune response. Using a transgenic fly system (SINrep) expressing a self-replicating viral genome, we have previously demonstrated the antiviral response of the Drosophila Imd (Immune Deficiency) and Jak-STAT innate immunity pathways. In the current study, microarray analysis of SINrep flies in comparison to control GFP flies aims to detect genes that are sensitive to Sindbis viral RNA replication.
Project description:A spectral library was built for Drosophila melanogaster. The spectral library allows reproducible quantification for thousands of peptides per SWATH-MS analysis.
Proteins from Drosophila melanogaster embryo, adult flies were digested with trypsin using in-gel digestion and the peptides were fractionated by high-pH reverse phase chromatography. HRM peptides were spiked into the peptides mixture and each fraction was injected on a Sciex TripleTOF 6600 mass spectrometer fitted with microflow set-up.
The resulting .wiff files were analysed using MaxQuant and Spectronaut.
