Novel Fluorescent Proteins for Bioimaging

描述(由申请人提供)： 来自水杉的绿色荧光蛋白(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。我们最近开始了一次探险，从世界这个地区收集荧光海洋标本。我们已经收集了大约100个样本并对其进行了部分表征，我们已经从大堡礁珊瑚中克隆了第一个新的荧光蛋白。我们正在申请申请(RFA：EB-03-003)“细胞和分子成像系统和方法的研究与开发”，继续这项工作，并克隆和表征新的荧光蛋白，用作生物成像研究的工具。我们的目标是：(1)构建和筛选从大堡礁采集的海洋生物新的荧光蛋白的cDNA文库；(2)从以下方面鉴定克隆的重组荧光蛋白：(A)光谱性质；(B)化学稳定性；(C)在细菌和培养的哺乳动物细胞中表达；(D)在小型动物非洲爪哇(Xenopus Laevis)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。

DESCRIPTION (provided by applicant): Green fluorescent protein from Aequoria victoria (GFP) 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. We recently embarked on an expedition to collect fluorescent marine specimens from this region of the world. About a hundred specimens have been collected and partially characterized, and we have cloned the first novel fluorescent protein from coral derived from the Great Barrier Reef. We are applying in response to the Request for Applications (RFA: EB-03-003) "Research and Development of Systems and Methods for Cellular and Molecular Imaging", to continue this work, and to clone and characterize novel fluorescent proteins for use as tools in bioimaging research. Our objectives are to: (1) Construct and screen cDNA libraries for novel fluorescent proteins from marine organisms collected from the Great Barrier Reef; and (2) Characterize cloned, recombinant fluorescent proteins with respect to: (a) spectral properties; (b) chemical stability; (c) expression in bacteria and cultured mammalian cells; (d) expression in a prototypic small animal, Xenopus laevis. 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.