Patent ID: 7666606
Filing Date: 2010-02-23
Classification: C40B,G01N

Abstract:
1. An assay for detecting the interaction of two proteins, X and Y, comprising: (a) expressing in a cell a first polynucleotide construct comprising the coding sequence of a soluble first microdomain tag fragment of a fluorescent protein, wherein the microdomain tag fragment corresponds to one or more beta-strands of the fluorescent protein, fused to the coding sequence of protein X; (b) expressing in the cell a second polynucleotide construct comprising the coding sequence of a soluble second microdomain tag fragment of the fluorescent protein, wherein the microdomain tag fragment corresponds to one or more other beta-strands of the fluorescent protein, fused to the coding sequence of protein Y, or introducing the fusion protein, Y-microdomain tag, encoded thereby into the cell; (c) expressing in the cell a third polynucleotide construct comprising the coding sequence of a soluble assay fragment complementary to the microdomain tag fragments of (a) and (b), wherein the assay fragment corresponds to the remaining beta-strands of the fluorescent protein, or introducing the assay fragment into the cell; (d) detecting fluorescence in the cell and thereby detecting the interaction of proteins X and Y, wherein the two microdomain tag fragments selected do not self-complement with the assay fragment in the absence of interacting proteins fused thereto.