Transgenic animals expressing artificial epitope-tagged proteins
AssigneeThe Regents of the University of California
InventorsStanley B. Prusiner, Glenn C. Telling, Fred E. Cohen, Michael R. Scott
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Abstract
DNA constructs are provided of epitope-tagged proteins or protein fragments which are conveniently purified with immunoaffinity chromatography such as epitope-tagged prion proteins (PrP). Transgenic animals expressing an epitope-tagged protein are provided, including transgenic animals expressing epitope-tagged PrP. Methods for distinguishing between the conformational shapes of a protein and a convenient method for isolating a tagged protein by immunoaffinity chromatographic methods are provided.
Claims2
A method for distinguishing between the conformational shapes of a prion protein having a first and second conformation shape, comprising the steps of:
- a) generating a recombinant nucleic acid construct comprising a nucleic acid sequence encoding an amino acid sequence encoding biologically active prion protein fragment and a heterologous epitope domain;
- b) transfecting a cell or organism with the construct of a);
- c) expressing said construct to produce an epitope-tagged prion protein, wherein said epitope is unexposed on the surface of said prion protein having said first conformational shape, and wherein said epitope is exposed on the surface of said prion protein having said second conformational shape;
- and
- d) distinguishing between said first and second conformational shapes by detecting the presence or absence of said epitope on the surface of said prion protein.
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