GFP-like proteins:Diversity and determinants of color

类 GFP 蛋白质：多样性和颜色的决定因素

• 批准号: 6797646
• 负责人: MIKHAIL V MATZ
• 金额: $ 2.19万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-16 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6797646
• 关键词: Anthozoa; SDS polyacrylamide gel electrophoresis; aminoacid; bioluminescence; biophysics; coral; fluorescence; green fluorescent proteins; polymerase chain reaction; protein purification; protein structure; proteins; sample collection; site directed mutagenesis

DESCRIPTION (provided by applicant): Proteins homologous to green fluorescent protein (GFP) from the jellyfish Aequorea victoria have become indispensable tools for in vivo imaging, and are widely used in basic biomedical research and drug discovery. Finding such proteins in non-bioluminescent Anthozoa offers an exciting opportunity to expand the technology by providing more colors to choose from. In addition, a number of novel features valuable for the engineering of advanced reporter systems have become available, such as precisely time-dependent changes of emission color or light-driven fluorescence activation/inhibition. However, since only a few limited-scale attempts have been undertaken thus far to clone GFP-like proteins from Anthozoa, it can be expected that many more proteins with novel spectral and structural features can be discovered during systematic survey of these organisms. In particular, these may include variants of existing proteins that are free of their drawbacks, such as oligomerization or slow chromophore maturation. Here we propose such a systematic study aimed at complete characterization of a diversity of GFP-like proteins in Anthozoa in order to create a large inventory of potential reporters for biomedical imaging applications. We intend to use the accumulated body of information on sequence/spectral features to pinpoint sequence determinants of coloration, and to verify their role by means of site-specific mutagenesis. This would greatly facilitate rational design of GFP-like reporters.

描述(申请人提供)：与水母绿色荧光蛋白(GFP)同源的蛋白质已成为体内成像不可缺少的工具，并广泛应用于基础生物医学研究和药物开发。在非生物发光的珊瑚虫中发现这样的蛋白质提供了一个令人兴奋的机会，通过提供更多的颜色可供选择来扩展这项技术。此外，许多对先进报告系统的工程有价值的新特征已经变得可用，例如精确地随时间改变发射颜色或光驱动的荧光激活/抑制。然而，由于到目前为止从珊瑚虫中克隆GFP样蛋白的尝试还很少，可以预期在对这些生物的系统调查中会发现更多具有新的光谱和结构特征的蛋白质。特别是，这些可能包括现有蛋白质的变种，没有它们的缺点，如齐聚或发色团成熟缓慢。在这里，我们提出了这样一项系统的研究，旨在完整地表征安东虫中的GFP样蛋白的多样性，以便为生物医学成像应用创建大量的潜在报告。我们打算利用积累的关于序列/光谱特征的信息来精确定位着色的序列决定因素，并通过定点突变的方法来验证它们的作用。这将极大地促进GFP类记者的合理设计。