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Reverse screening and target identification with minicells

  • US 20030224369A1
  • Filed: 05/28/2002
  • Published: 12/04/2003
  • Est. Priority Date: 02/25/2002
  • Status: Abandoned Application
First Claim
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1. A method of identifying a nucleic acid that encodes a protein that binds to or chemically alters a preselected ligand, comprising:

  • (a) separately contacting said ligand with individual members of a minicell library, wherein minicells in said library comprise expression elements, wherein said expression elements comprise DNA inserts, wherein an ORF in said DNA insert is operably linked to expression sequences, in order to generate a series of reaction mixes, each reaction mix comprising a different member of said minicell library;

    (b) incubating said reaction mixes, thereby allowing a protein that binds to or chemically alters said preselected ligand to bind or chemically alter said preselected ligand;

    (c) detecting a change in a signal from reaction mixes in which said ligand has been bound or chemically altered;

    (d) preparing DNA from reaction mixes in which said ligand has been bound or chemically altered;

    wherein said DNA is a nucleic acid that encodes a protein that binds to or chemically alters said preselected ligand.

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