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: 6507916
• 负责人: MIKHAIL V MATZ
• 金额: $ 24.25万
• 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/6507916
• 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（由申请人提供）：与来自水母Aequorea维多利亚的绿色荧光蛋白（GFP）同源的蛋白质已成为体内成像不可或缺的工具，并广泛用于基础生物医学研究和药物发现。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该技术。此外，许多新的功能，有价值的先进的报告系统的工程已经成为可用的，如精确的时间依赖性变化的发射颜色或光驱动的荧光激活/抑制。然而，由于迄今为止仅进行了一些有限规模的尝试来从珊瑚虫中克隆GFP样蛋白，因此可以预期，在这些生物体的系统调查期间可以发现更多具有新的光谱和结构特征的蛋白质。特别地，这些可以包括现有蛋白质的变体，这些变体没有它们的缺点，例如寡聚化或缓慢的发色团成熟。在这里，我们提出了这样一个系统的研究，目的是完整的表征珊瑚虫中的GFP样蛋白的多样性，以创建一个大的库存的潜在记者的生物医学成像应用。我们打算使用积累的信息体的序列/光谱特征，以查明着色的序列决定因素，并验证其作用，通过定点诱变。这将极大地促进GFP样报告基因的合理设计。