Project description:Detection of species-specific proteotypic peptides for accurate and easy characterization of infectious non-tuberculous mycobacteria such as Mycobacterium avium subsp. paratuberculosis is essential. Therefore, we conducted an in-depth global proteomic experiment using M. avium subsp. paratuberculosis ATCC 19698 (Map) strain followed by proteome database search and spectral library generation. The lysate was subjected to in-solution proteomic sample preparation and fractionated using an offline C18 StageTip. Each fraction was acquired in technical triplicates using a 180 min data-dependent acquisition (DDA) method in Orbitrap Fusion Tribrid (Thermo Scientific) mass spectrometer. The resulting raw DDA data were searched against the M. avium subsp. paratuberculosis proteome database using Proteome Discoverer and FragPipe. The resulting peptide spectrum matches were converted into a spectral library using BiblioSpec.
Project description:Comparsion of proteomes of Campylobacter fetus subsp. fetus to compare protein level via iBAQ analysis, expression (by LFQ) and coverage between Campylobacter fetus subsp. fetus strain82-40 vs Campylobacter fetus subsp. fetus strain ATCC 27374
Project description:Background: Pantoea ananatis LMG 2665T synthesizes and utilizes acyl homoserine lactones (AHLs) for signaling. In this strain, short chain AHLs (C4 to C8) are produced by the EanI/R quorum sensing (QS) system that is involved in pathogenicity and biofilm formation. The complete set of genes regulated by the EanI/R system in P. ananatis LMG 2665T is still not fully known. In the present study, RNA-seq was used to analyze the transcriptome profiles controlled by the EanI/R system in this strain by comparing the wild type strain and its QS mutant 2665T ean∆I/R during lag and log stages. The RNA seq data was validated by RT qPCR. Results: The results showed that the EanI/R regulon in P. ananatis LMG 2665T comprised 144 genes, constituting 3.3% of the whole transcriptome under the experimental conditions in this study. The majority of genes regulated by the EanI/R system included genes for flagella assembly, bacterial chemotaxis, pyruvate metabolism, two component system, metabolic pathways, microbial metabolism and biosynthesis of secondary metabolites. Conclusions: This is the first study to identify the EanI/R QS regulon in P. ananatis LMG 2665T. Functional analysis of genes regulated the EanI/R system in LMG 2665T could help unveil genes that play a vital role in pathogenesis and survival strategies of this pathogen.