Patent ID: 6905818
Filing Date: 2005-06-14
Classification: C12N,G01N

Abstract:
1. A method for reducing false positives in the identification of at least one member of a pair or complex of interacting molecules from potentially interacting molecules, comprising: (A) providing at least one set of host cells, each set containing at least one genetic element comprising a selectable marker, said selectable marker being different between different sets of host cells, said genetic element comprising a nucleic acid encoding one of said potentially interacting molecules, said host cells further carrying a readout system that is activated upon the presence of auto-activating molecules; (B) transferring at least one set of said host cells or progeny of at least one set of said host cells to at least one selective medium, different for each set of host cells, which allows growth of said host cells in the presence of said genetic element comprising a selectable marker and which precludes growth of said host cells upon auto-activation of said readout system, thereby selecting against host cells expressing a molecule able to auto-activate the readout system; (C) combining in said host cells at least two said genetic elements, wherein at least one set of said host cells with one of said at least two genetic elements undergoes the selecting step as specified in (B); (D) allowing at least one interaction between said potentially interacting molecules, if any, to occur; (E) selecting for said interaction by transferring said host cells or progeny of said host cells to a selective medium that allows identification of said host cells upon activation of the readout system; (F) identifying host cells that contain interacting molecules that activate said readout system on said selective medium; (G) identifying at least one member of said pair or complex of interacting molecules; wherein said host cells are not yeast cells.