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

RIG：印度-太平洋和加勒比海海洋生物荧光蛋白的分离、表征和进化

• 批准号: 0920572
• 负责人: David Gruber
• 金额: $ 19.93万
• 依托单位: CUNY Baruch College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0920572&HistoricalAwards=false
• 关键词: RIG; Isolation; Characterization; Evolution; Fluorescent

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.

来自Aequoria Victoria的智能绿色荧光蛋白(GFP)使细胞生物成像研究发生了革命性的变化。GFP已被用于病毒、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。绿色荧光蛋白的使用刺激了大量的突变研究，以增强绿色荧光蛋白的有效性，并试图探索来自Aequoria以外的物种的荧光蛋白。对于许多生物成像应用，可用的荧光蛋白质库仍然是有限的。到目前为止，蕴藏荧光的海洋生物种类最多的是位于澳大利亚东海岸外的大堡礁。该项目的首席调查员(PI)最近共同领导了一支从世界这一区域收集荧光海洋标本的探险队。大约100个珊瑚和海葵标本已经被收集、鉴定，现在被低温储存在纽约市的美国自然历史博物馆(AMNH)。PI将在巴鲁克学院的纽约市立大学(CUNY)建立这一领域的研究计划，并进行克隆和表征新的荧光蛋白，作为生物成像研究的工具。研究目标是：(1)构建和筛选从澳大利亚大堡礁采集的海洋生物新的荧光蛋白的cDNA文库，并在加勒比海最原始的珊瑚礁之一小开曼岛进行交叉比较研究；(2)从以下方面鉴定克隆的重组荧光蛋白：(A)光谱特性；(B)化学稳定性；(C)在细菌细胞中表达。目前对这种新工具的需求很大，预计通过该项目表征的新的荧光蛋白将在医学和生物学的许多分支中获得广泛的应用。更广泛的影响这个项目旨在培养和激励纽约州立大学巴鲁克学院(CUNY Baruch College)(经常被认为是美国种族最多元化的校园)的本科生开始生物职业生涯。学生群体反映了纽约市的多样性，超过50%的学生是家里第一个上大学的人。国际学生联合会将集中招收和指导将在实地和实验室生态研究方面获得经验的少数族裔学生。巴鲁克学院、美国自然历史博物馆、耶鲁大学和布朗大学将建立伙伴关系，少数族裔本科生将接触到多样化的生物学研究，并将有机会在合作者的实验室中获得额外的指导。作为巴鲁克生物医学学会的导师，PI将吸收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向公众宣传外，PI还计划在AMNH的科学简报网站上发布研究结果。科学简报是一个创新的在线和展览项目，为公众提供了一个了解科学发现的兴奋之窗。它面向6到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。