Patent ID: 9188592
Filing Date: 2015-11-17
Classification: G01N,Y10T

Abstract:
1. A method of detecting the folding of a protein domain, the method comprising: performing cell free translation on a nucleic acid construct in the presence of a ribosome, the construct comprising a first polynucleotide that encodes the protein domain, a second polynucleotide that encodes a protein tag, and a third polynucleotide that encodes a FRET donor, the FRET donor having an emission wavelength, thereby generating a ribosome-nascent chain complex comprising the ribosome, the protein domain, the protein tag, and the FRET donor; incorporating a FRET acceptor into the protein domain; purifying the ribosome-nascent chain complex; conjugating the ribosome-nascent chain complex to a solid substrate via the tag; detecting a first fluorescence signal at the emission wavelength of the FRET donor, wherein a fluorescence signal that is lower than the fluorescence signal of a protein domain with a FRET donor but lacking a FRET acceptor indicates folding of the protein domain.