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

来自维多利亚水母的绿色荧光蛋白（GFP）彻底改变了细胞生物成像研究。GFP已用于病毒、细菌、培养细胞、酵母、转基因植物和从蠕虫到小鼠的转基因动物。GFP的效用已经刺激了许多突变研究以增强GFP的有用性，以及尝试探索来自水母属以外的物种的荧光蛋白。对于许多生物成像应用，荧光蛋白的可用库仍然有限。到目前为止，拥有荧光的海洋生物的最大多样性存在于位于澳大利亚东海岸的大堡礁。该项目的首席研究员（PI）最近共同领导了一个探险队，从世界这一地区收集荧光海洋标本。大约有100个珊瑚和海葵标本已经被收集起来，并进行了鉴定，目前存放在纽约市的美国自然历史博物馆（AMNH）的低温储存中。 PI将在纽约（CUNY）巴鲁克学院建立这一领域的研究计划，并进行克隆和表征新型荧光蛋白，用作生物成像研究的工具。 研究目标是：（1）建立和筛选从澳大利亚大堡礁收集的海洋生物的新荧光蛋白的cDNA文库，并着手对加勒比最原始的珊瑚礁之一小开曼岛进行交叉比较研究;（2）鉴定克隆的重组荧光蛋白的下列特性：（a）光谱特性;（B）化学稳定性;和（c）在细菌细胞中表达。目前对这种新工具有很大的需求，并且预计从该项目中表征的新荧光蛋白将在医学和生物学的许多分支中找到广泛的适用性。更广泛的影响这个项目旨在培养和激励来自纽约市立巴鲁克学院（经常被认为是全国种族最多样化的校园）的本科生从事生物学职业。学生群体反映了纽约市的多样性，超过50%的学生是家庭中第一个上大学的。PI将集中招募和指导少数民族学生，这些学生将获得实地和实验室生态研究的经验。巴鲁克学院、美国自然历史博物馆、耶鲁大学和布朗大学之间将建立伙伴关系，其中少数民族本科生将接触到生物研究的多样性，并将有机会在合作者的实验室获得额外的指导。作为巴鲁克生物医学学会的导师，PI将利用他的生物课和学会的学生，并将研究结果纳入他的教学课程。除了作为一名科学记者进行公众宣传外，PI还计划在AMNH的科学公报网站上发表研究结果。 Science Bulletins是一个创新的在线和展览计划，为公众提供了一个了解科学发现兴奋的窗口。它面向6至12年级的学生和他们的老师，去年有超过25万的独立访问者。

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.