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Chimeric green fluorescent protein-aequorin as bioluminescent Ca2+ reporters at the single-cell level

Baubet, V., Le Mouellic, H., Campbell, Anthony Keith, Lucas-Meunier, E., Fossier, P. and Brulet, P. 2000. Chimeric green fluorescent protein-aequorin as bioluminescent Ca2+ reporters at the single-cell level. Proceedings of the National Academy of Sciences of the United States of America 97 (13) , pp. 7260-7265. 10.1073/pnas.97.13.7260

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Monitoring calcium fluxes in real time could help to understand the development, the plasticity, and the functioning of the central nervous system. In jellyfish, the chemiluminescent calcium binding aequorin protein is associated with the green fluorescent protein and a green bioluminescent signal is emitted upon Ca2+ stimulation. We decided to use this chemiluminescence resonance energy transfer between the two molecules. Calcium-sensitive bioluminescent reporter genes have been constructed by fusing green fluorescent protein and aequorin, resulting in much more light being emitted. Chemiluminescent and fluorescent activities of these fusion proteins have been assessed in mammalian cells. Cytosolic Ca2+ increases were imaged at the single-cell level with a cooled intensified charge-coupled device camera. This bifunctional reporter gene should allow the investigation of calcium activities in neuronal networks and in specific subcellular compartments in transgenic animals.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Subjects: Q Science > QD Chemistry
R Medicine > RM Therapeutics. Pharmacology
Publisher: National Academy of Sciences
ISSN: 0027-8424
Last Modified: 05 Jun 2017 02:58

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