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.

来自维多利亚州的Equoria的智力精美荧光蛋白（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的效用刺激了许多突变研究，以增强GFP的实用性，并尝试探索来自Aquoria以外的物种的荧光蛋白。对于许多生物成像应用，荧光蛋白的可用曲目仍然有限。到目前为止，位于澳大利亚东海岸附近的大堡礁上，海洋生物的最大多样性存在着荧光。该项目的首席研究员（PI）最近共同领导了一次从这个地区收集荧光海洋标本的探险。已经收集了大约一百个珊瑚和海葵标本，并在纽约市的美国自然历史博物馆（AMNH）中进行了寒冷的存储。 PI将在纽约市城市大学（CUNY）的该地区建立一项研究计划，并进行克隆并表征新型荧光蛋白，以用作生物成像研究的工具。 研究目标是：（1）从澳大利亚大堡礁收集的海洋生物的新型荧光蛋白的构造和筛选cDNA库，并在加勒比海最原始的珊瑚礁之一的小开曼岛（Little Cayman Island）上进行了一项交叉比较研究； （2）与以下方面的克隆，重组荧光蛋白的特征：（a）光谱特性； （b）化学稳定性； （c）在细菌细胞中的表达。目前，这种新工具非常需要，预计以该项目为特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。更广泛的影响力旨在培训和激发来自Cuny Baruch College（经常被认为是全国最多样化的校园）的本科生，以开始生物学的职业。学生团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人。 PI将深入招募和导师的少数族裔学生，这些学生将在基于实地和实验室的生态研究中获得经验。伙伴关系将在巴鲁克学院，美国自然历史博物馆，耶鲁大学和布朗大学之间建立，其中少数本科生将受到生物学研究的多样性，并有机会在合作者实验室中进行额外的指导。作为Baruch生物医学协会的导师，PI将吸引他的生物学课程和社会的学生，并将发现纳入他的教学课程中。除了他作为科学记者的公开宣传外，PI还计划在AMNH的科学公告网站上发布调查结果。 科学公告是一项创新的在线和展览计划，为公众提供了科学发现兴奋的窗口。它针对6至12年级的学生及其老师，去年有超过25万个唯一访客的观众。