×

Identification and characterization of interacting molecules

  • US 6,664,048 B1
  • Filed: 05/30/2000
  • Issued: 12/16/2003
  • Est. Priority Date: 11/27/1997
  • Status: Expired due to Fees
First Claim
Patent Images

1. A method for detecting formation of complexes including a first test member and a potentially interacting second test member, comprising:

  • (a) providing host cells containing;

    (i) a first type of genetic element associated with at least one each of a selectable and a counter-selectable marker, said first type of genetic element comprising nucleotide sequences encoding one of said first test member(s), and one of two parts of a functional entity;

    (ii) a second type of genetic element associated with at least one each of a selectable and a counter-selectable marker, said second type of genetic element comprising nucleotide sequences encoding one of said second test member(s), and the other of two parts of said functional entity;

    wherein selectable and counter-selectable marker(s) associated with the first and second types of genetic elements differ by at least one selectable and at least one counter-selectable marker(s), respectively; and

    (iii) a readout system for producing a detectable signal;

    wherein interaction between said first and second test members brings said two parts of the functional entity into close proximity, thereby activating said readout system and producing the detectable signal;

    (b) culturing the host cells under conditions wherein complexes including said first and second test members, if any, will be formed in the cell;

    (c) selecting host cells wherein at least one of said complexes has formed by transferring samples of said host cells to;

    (i) at least two different selective media, wherein each of said selective media allows growth of said host cells only in the absence of said counter-selectable marker(s) associated with one of said two types of genetic elements and in the presence of said selectable marker(s) associated with the other of said two types of genetic elements, wherein at least one of said at least two selective media selects for the presence of said first genetic element and the absence of said second genetic element, and at least a second of said at least two selective media selects for the presence of said second genetic element and the absence of said first genetic element; and

    (ii) a further selective medium that allows identification of said host cells only on the activation of said readout system; and

    (d) detecting formation of at least one of said complexes by identifying host cells wherein;

    (i) said readout system is not activated on any of said selective media specified in step (c)(i);

    but which (ii) said readout system is activated on said selective medium specified in step (c)(ii).

View all claims
  • 1 Assignment
Timeline View
Assignment View
    ×
    ×