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Using MS2 affinity pulldown to identify sRNA associating proteins in the bacterium Deinococcus radiodurans

ABSTRACT: We carried out the widely used MS2 aptamer approach to identify the proteins that could potentially interact with sRNAs in D. radiodurans. The MS2 aptamer was fused to the 5 extremity of PprS/Dsr2, a well-characterized D. radiodurans sRNA . This construct is then expressed off the pRADgro plasmid in wild-type D. radiodurans at mid-exponential phase. After bacterial lysis, the crude extract was loaded into an amylose-based chromatography column previously coated with the MS2 protein fused to the maltose-binding protein. This enables the specific capture of MS2-Dsr2 and its interacting protein partners. After elution, co-purified proteins were identified by LC-MS/MS and subsequent bioinformatic analysis

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Deinococcus Radiodurans R1 (ncbitaxon:243230)

SUBMITTER: Lydia M. Contreras

PROVIDER: MSV000094854 | MassIVE | Thu May 23 13:39:00 BST 2024

SECONDARY ACCESSION(S): PXD052534

REPOSITORIES: MassIVE

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