Fusion proteins useful for detecting analytes
First Claim
1. A fusion protein comprising:
- (a) two detection portions, wherein (i) the first detection portion is an energy-emitting protein portion and the second detection portion is a fluorescent protein portion;
or (ii) the two detection portions are portions of a split fluorescent protein; and
(b) a periplasmic binding protein (PBP) portion between said two detection portions, which undergoes a conformational change upon binding of a compound;
wherein said conformational change results in a change of the energy emitted by said two detection portions.
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Accused Products
Abstract
Disclosed are fusion proteins comprising two detection portions and a periplasmic binding protein (PBP) portion between this two detection portions which undergoes a conformational change upon binding of a compound. Also described are nucleic acid molecules comprising a nucleotide sequence encoding such a fusion protein as well as expression cassettes, vectors, transgenic plants and transgenic non-human animals comprising said nucleic acid molecules. In addition, described are methods for detecting an analyte in a sample or in a cell using said fusion protein, and optionally, a control sensor allowing a calibration of analyte detection made by said fusion protein. Also described are such control sensors and nucleic acid molecules encoding them, as well as diagnostic compositions, kits and uses of the fusion protein and the control sensor for in vitro or in vivo analyte detection and preparing a diagnostic composition.
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Citations
46 Claims
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1. A fusion protein comprising:
- (a) two detection portions, wherein (i) the first detection portion is an energy-emitting protein portion and the second detection portion is a fluorescent protein portion;
or (ii) the two detection portions are portions of a split fluorescent protein; and
(b) a periplasmic binding protein (PBP) portion between said two detection portions, which undergoes a conformational change upon binding of a compound;
wherein said conformational change results in a change of the energy emitted by said two detection portions. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 41, 42, 43, 44, 45)
- (a) two detection portions, wherein (i) the first detection portion is an energy-emitting protein portion and the second detection portion is a fluorescent protein portion;
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36. A control sensor comprising:
- (a) two detection portions, whereby (i) the first detection portion is an energy-emitting protein portion and the second detection portion is a fluorescent protein potion;
or (ii) the two detection portions are portions of a split fluorescent protein and (b) a PBP portion fused to said first and second detection portion which is modified with respect to the corresponding wild-type PBP portion and therefore is incapable of binding the compound which said wild-type PBP portion binds;
wherein said first and/or second detection portion emits energy. - View Dependent Claims (37, 38, 39, 46)
- (a) two detection portions, whereby (i) the first detection portion is an energy-emitting protein portion and the second detection portion is a fluorescent protein potion;
Specification