Recombinant phycobiliproteins with enhanced fluorescence and photochemical properties
AssigneesBoard of Supervisors of the University of Louisiana System, The Penn State Research Foundation
InventorsDonald A. Bryant, Wendy M. Schluchter, Richard M. Alvey, Avijit Biswas
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Abstract
Novel fluorescent phycobiliprotein fusion proteins and methods of use are described. The novel phycobiliproteins can be produced in a cell comprising two or more heterodimeric lyases, an apoprotein and a bilin reductase, which components react to form the phycobiliprotein fusion protein. Also described are phycobiliprotein based transcription reporter cells and assays, which cells conditionally express a heterologous, fluorescent, phycobiliprotein domain even in anoxic conditions.
Claims36
A modified phycobiliprotein selected from the group consisting of:
- HT-CpcA-PΦB, HT-CpcA-PEB, HT-CpcA-PCB, HT-CpcA-PtVB, HT-CpcA-PVB, and HT-CpcA-PUB, HT-ApcF-PEB, HT-ApcA-PEB, HT-ApcF-PΦB, HT-ApcB-PEB, HT-ApcB-PΦB, HT-ApcD-PEB, HT-ApcD-PΦB.
A modified phycobiliprotein with a PCB chromophore attached to CpcA wherein said phycobiliprotein has an absorption maximum of 625, a fluorescence emission maximum of 646, and a fluorescence quantum yield of 0.39.
A modified phycobiliprotein with a PEB chromophore attached to CpcA wherein said phycobiliprotein has an absorption maximum of 556, a fluorescence emission maximum of 568, and a fluorescence quantum yield of 0.98.
A modified phycobiliprotein with a PΦB chromophore attached to CpcA wherein said phycobiliprotein has an absorption maximum of 637, a fluorescence emission maximum of 656, and a fluorescence quantum yield of 0.18.
A modified phycobiliprotein with a PVB chromophore attached to CpcA wherein said phycobiliprotein has an absorption maximum of 561, a fluorescence emission maximum of 577, and a fluorescence quantum yield of 0.14.
A modified phycobiliprotein with a PUB chromophore attached to CpcA wherein said phycobiliprotein has an absorption maximum of 497.
A modified phycobiliprotein with a PtVB chromophore attached to CpcA wherein said phycobiliprotein has an absorption maximum of 575, a fluorescence emission maximum of 590, and a fluorescence quantum yield of 0.23.
A recombinant cell which expresses a phycobiliprotein fusion protein comprising:
- two or more heterodimeric lyases, an apoprotein and a bilin reductase, which components react inside the cell to form the phycobiliprotein protein in the presence of biliverdin IXα.
A novel bilin lyase TeCpcS having an amino acid sequence of SEQ ID NO:
- 24.
A novel bilin lyase TcCpcS encoded by SEQ ID NO:25
Dependencies are inferred from the claim text.