Project description:Using recent developments in sample preparation strategies and improvements in mass spectrometry (MS), an optimized procedure was developed to characterize the proteome of Methylocystis sp. strain SC2, a type II methanotroph. It represents one of the ecologically important groups of methane-oxidizing bacteria. The major challenge for developing an efficient analytical proteomics workflow for methanotrophic bacteria is the high amount of membrane-associated proteins that need to be efficiently solubilized and digested for downstream analysis. Therefore, each step of the workflow, including cell lysis, protein solubilization and digestion, and MS peptide quantification, was assessed and optimized. Our novel crude-lysate-MS approach proved to increase protein quantification accuracy and the proteome coverage of strain SC2. It captured 62% of predicted SC2 proteome, with 10-fold increase in membrane-associated proteins relative to less effective conditions. Use of crude cell lysate for downstream analysis showed not only to be highly efficient for strain SC2 but also for other members of the Methylocystaceae family. To validate the efficiency of our newly developed workflow, we analyzed the SC2 proteome under two contrasting nitrogen conditions, with a focus on the differential expression of proteins involved in methane and nitrogen metabolisms.

2018-07-23 | PXD009027 | Pride

Genome-wide binding site profiling of NdgR in Streptomyces coelicolor

Project description:We identified genome-wide binding regions of NdgR in Streptomyces coelicolor using chromatin immunoprecipitation sequencing (ChIP-seq). We constructed 6×myc-tagged NdgR strain using homologous recombination with myc-tagging vector. Analysis of the sequencing data aligned to Streptomyces coelicolor genome database (NC_003888).

2014-12-29 | GSE59010 | GEO

Pulveribacter suum strain:SC2-7

Project description:Pulveribacter suum strain:SC2-7 Genome sequencing

| PRJNA438161 | ENA

Chryseobacterium suipulveris strain:SC2-2

Project description:Chryseobacterium suipulveris strain:SC2-2 Genome sequencing

| PRJNA820572 | ENA

Paracoccus suum strain:SC2-6

Project description:Paracoccus suum strain:SC2-6 Genome sequencing

| PRJNA478802 | ENA