Detection of in vivo Protein Interactions in All Bacterial Components by Forster Resonance Energy Transfer with the Superfolder mTurquoise2 ox-mNeongreen FRET Pair.

ABSTRACT: This protocol was developed to qualitatively and quantitatively detect protein-protein interactions in all compartments of Escherichia coli by Förster Resonance Energy Transfer (FRET) using the Superfolder mTurquoise2 ox-mNeonGreen FRET pair (sfTq2^ox-mNG). This FRET pair has more than twice the detection range for FRET interaction studies in the cytoplasm or periplasm of E. coli compared to other pairs to date. These protein-interaction studies can be performed in vivo because fluorescent proteins can be genetically encoded as fusions to proteins of interest and expressed in the cell. sfTq2^ox and mNG fluorescent protein fusions are co-expressed in bacterial cells and the fluorescence emission spectra are measured. By also measuring reference spect

SUBMITTER: Meiresonne NY 

PROVIDER: S-EPMC7853951 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature