Detection of Protein Interactions in the Cytoplasm and Periplasm ofEscherichia coli by Forster Resonance Energy Transfer.

ABSTRACT: This protocol was developed to qualitatively and quantitatively detect protein-protein interactions in Escherichia coli by Förster Resonance Energy Transfer (FRET). The described assay allows for the previously impossible in vivo screening of periplasmic protein-protein interactions. In FRET, excitation of a donor fluorescent molecule results in the transfer of energy to an acceptor fluorescent molecule, which will then emit light if the distance between them is within the 1-10 nm range. Fluorescent proteins can be genetically encoded as fusions to proteins of interest and expressed in the cell and therefore FRET protein-protein interaction experiments can be performed in vivo. Donor and acceptor fluorescent protein fusions are constructed for bacterial proteins that a

SUBMITTER: Meiresonne NY 

PROVIDER: S-EPMC8203949 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature