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US 2006/0223089 A1Published application
Reverse two-hybrid systems
AssigneesThe Johns Hopkins University, The General Hospital Corporation
InventorsMarc Vidal, Jef Boeke, Ed Harlow
Life of this patent
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Abstract
Disclosed are methods for identifying molecular interactions (e.g., protein/protein, protein/DNA, protein/RNA, or RNA/RNA interactions). All of the methods within the invention employ counterselection and at least two hybrid molecules. Molecules which interact reconstitute a transcription factor and direct expression of a reporter gene, the expression of which is then assayed. Also disclosed are genetic constructs which are useful in practicing the methods of the invention.
Claims14
1-107. (canceled)
A cell comprising:
- a) a counterselectable reporter gene operably linked to a first DNA binding protein recognition site;
- b) a first fusion gene which expresses a first hybrid protein, said first hybrid protein comprising a test protein covalently bonded to a DNA binding moiety which specifically binds to said DNA binding protein recognition site;
- and
- c) a second fusion gene which expresses a second hybrid protein, said second hybrid protein comprising a test protein covalently bonded to a gene activating moiety.
1 more element
A cell comprising:
- i) a first reporter gene operably linked to a first DNA binding protein recognition site;
- and
- ii) a second reporter gene operably linked to a second DNA binding protein recognition site;
- wherein said first and said second DNA binding protein recognition sites are different.
1 more element
Dependencies are inferred from the claim text.