Project description:Pandoravirus aubagnensis was isolated from freshwater collected from “Manon des Sources” in Marseille city,France. By co-culture on Acanthamoeba castellanii. Pandoravirus belohorizontensis was isolated from soil samples collected from Belo Horizonte city in Brazil, by co-culture on Acanthamoeba castellanii.
Project description:Proteome analysis of a novel type of virus. The virus was grown in A. castellanii amobea before being purified. Viral particules were enriched after centrifugation and proteins solubilised by SDS before being stacked in the top of a SDS-PGE gel. After in-gel digestion, resulting peptides were injected for a 120min nanoLC-MS/MS analysis using an Ultimate U3000 system and a LTQ-Orbitrap Velos pro hybrid mass spectrometer (Top 20).Data processing and bioinformatics: Data were processed automatically using Mascot Daemon software (version 2.3.2, Matrix Science). Concomitant searches against Pandoravirus and A. castellanii protein sequence databanks as well as classical contaminants database and the corresponding reversed databases were performed using Mascot (version 2.4). ESI-TRAP was chosen as the instrument, trypsin/P as the enzyme and 2 missed cleavage allowed. Precursor and fragment mass error tolerances were set respectively at 10 ppm and 0.6 Da. Peptide modifications allowed during the search were: carbamidomethyl (C, fixes) acetyl (N-ter, variable), oxidation (M, variable) and deamidation (NQ, variable). The IRMa software (Dupierris et al., Bioinformatics, 2009, 25:1980-1, version 1.30.4) was used to filter the results: selection of rank 1 peptides, peptide identification FDR < 1% (as calculated by employing the reverse database strategy), and minimum of 1 specific peptide per identified protein group.
Project description:Gymnocypris przewalskii przewalskii is distributed in Qinghai Lake, the largest inland saltwater lake in China. It is the only Cyprinidae fish in the Qinghai Lake water system and has extremely strong adaptability to the ecological environment with high salinity. G. p. przewalskii originates from the freshwater species Gymnocypris eckloni eckloni in the Yellow River and has a freshwater subspecies, Gymnocypris przewalskii ganzihonensis, distributed in the Ganzi River. Therefore, G. p. przewalskii is considered an ideal material for studying the high salt adaptation of plateau fish. Previous studies have characterized the evolutionary basis of highland adaptation in G. p. przewalskii; however, its adaptability to highly saline aquatic environments remains elusive. In the current study, we performed physiological, histological, genomic and transcriptomic analyses to investigate the phenotypical adaptation of G. p. przewalskii to a high saline environment and the underlying genomic and regulatory bases.