RIG: Isolation, Characterization and Evolution of Fluorescent Proteins from Indo-Pacific and Caribbean Marine Organisms

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Intellectual MeritGreen fluorescent protein (GFP) from Aequoria victoria has revolutionized cellular bioimaging studies. GFP has been used in viruses, bacteria, cultured cells, yeast, transgenic plants, and transgenic animals from worms to mice. The utility of GFP has stimulated numerous mutational studies to enhance the usefulness of GFP as well as attempts to explore fluorescent proteins from species other than Aequoria. The available repertoire of fluorescent proteins remains limited for many bioimaging applications. By far, the greatest diversity of marine life harboring fluorescence exists on the Great Barrier Reef, located off the eastern coast of Australia. The Principal Investigator (PI) on this project recently co-led an expedition to collect fluorescent marine specimens from this region of the world. About a hundred coral and anemone specimens have been collected, characterized, and are now in cryogenic storage at the American Museum of Natural History (AMNH) in New York City. The PI will establish a research program in this area at City University of New York (CUNY), Baruch College, and undertake cloning and characterizing novel fluorescent proteins for use as tools in bioimaging research. The research objectives are to: (1) Construct and screen cDNA libraries for novel fluorescent proteins from marine organisms collected from Australia's Great Barrier Reef and to embark on a cross-comparison study on Little Cayman Island, one of the Caribbean's most pristine coral reefs; and (2) Characterize cloned, recombinant fluorescent proteins with respect to: (a) spectral properties; (b) chemical stability; and (c) expression in bacteria cells. There is currently a great need for such new tools, and it is expected that the new fluorescent proteins characterized from this project will find wide applicability in many branches of medicine and biology. Broader ImpactThis project aims to train and inspire undergraduate students from CUNY Baruch College (frequently recognized as the most ethnically diverse campus in the nation) to embark on careers in biology. The student body reflects the diversity of NYC, and over 50% of the students are the first in their family to attend college. The PI will intensively recruit and mentor minority students that will gain experience in both field- and laboratory-based ecological research. Partnerships will be formed among Baruch College, The American Museum of Natural History, Yale University and Brown University in which minority undergraduate students will be exposed to a diversity of biological research and will have the opportunity for additional mentoring in the laboratories of collaborators. As a mentor to the Baruch BioMedical Society, the PI will draw upon students from his biology classes as well as the Society and incorporate findings into his teaching curriculum. In addition to his public outreach as a science journalist, the PI also plans to publish findings on AMNH's Science Bulletins website. Science Bulletins is an innovative online and exhibition program that offers the public a window into the excitement of scientific discovery. It is aimed toward 6th through 12th grade students and their teachers, and had a measured audience of over 250,000 unique visitors last year.