Patent ID: 6156511
Filing Date: 2000-12-05
Classification: C12N,C12Q

Abstract:
A method of isolating a DNA binding protein comprising:(a) providing a recombinant DNA vector comprising a coding sequence for a peptide having a specific affinity for a receptor in frame with a DNA segment encoding a linker;(b) inserting a library of oligonucleotides encoding different potential DNA binding proteins into multiple copies of the recombinant DNA vector in frame with the peptide coding sequence and the linker to form a library of different vectors encoding different fusion proteins, each comprising a potential DNA binding protein, a linker and the peptide, the fusion proteins differing in the potential DNA binding proteins;(c) transforming host cells with the library of different vectors to form transformed host cells;(d) culturing the transformed host cells under conditions suitable for expression of the fusion proteins, whereby, if a fusion protein comprises a potential DNA binding protein with affinity for a vector encoding the fusion protein, the fusion protein binds to the vector to form a complex;(e) lysing the transformed host cells under conditions such that complexes formed in (d) remain associated;(f) contacting the complexes with a receptor under conditions conducive to specific binding of the peptide to the receptor;(g) isolating complexes bound to the receptor, the complexes containing vectors encoding DNA binding proteins.