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1. (WO2017182437) QUANTIFICATION OF AN INTERACTION BETWEEN A RAS PROTEIN AND A RAF PROTEIN

Pub. No.:    WO/2017/182437    International Application No.:    PCT/EP2017/059123
Publication Date: Fri Oct 27 01:59:59 CEST 2017 International Filing Date: Wed Apr 19 01:59:59 CEST 2017
IPC: G01N 33/68
C12Q 1/66
Applicants: UNIVERSITÄT INNSBRUCK
Inventors: STEFAN, Eduard
BACHMANN, Verena
RAFFEINER, Philipp
RÖCK, Ruth
Title: QUANTIFICATION OF AN INTERACTION BETWEEN A RAS PROTEIN AND A RAF PROTEIN
Abstract:
The present invention provides a method for quantification of an interaction between a Ras protein and a Raf protein, comprising the steps of a) providing a cell expressing two constructs for a protein-fragment complementation assay, wherein the first construct comprises the Ras protein and a first luciferase protein fragment, wherein the first luciferase protein fragment is located C-terminal from the Ras protein and wherein the second construct comprises the Raf protein and a second luciferase protein fragment, wherein the second luciferase protein fragment is located C-terminal from the Raf protein, wherein the second construct comprises a full-length Raf sequence section, and wherein the first and the second luciferase protein fragments are derived from different sections of the same luciferase protein b) allowing the first and second constructs to contact each other, c) providing a bioluminescence substrate for an intact luciferase protein and d) measuring for bioluminescence, wherein said bioluminescence indicates interaction between the two constructs upon reassembling of the first and second luciferase protein fragments.