BIOSENSORS AND DYNAMICS OF GREEN FLUORESCENT PROTEIN

描述：(逐字摘自申请人的摘要)设计的生物传感器 通过不同种类的荧光变化来检测并提供可见指示 细胞内条件的研究将通过基因工程来实现 以水母中的绿色荧光蛋白为模板。 已经开发出了能够对pH和卤化物的变化做出反应的传感器 (氯)浓度，这是正常的细胞功能所必需的。 它们将在功能上得到改进，供细胞和分子生物学家在 单个活细胞，包括人类细胞，以便简化对细胞的研究 正常细胞和病变细胞中的突起。此外，还建议发展 通过可逆途径对氧化还原电位变化敏感的变体 用于检测氧化还原的二硫键的形成或断裂 亚细胞隔室，如内质网中的电位。 变体将通过吸光度和荧光光谱进行表征，并 潜在有用的生物传感器的结构将由X射线确定。 结晶学，以评估分子的结构基础 属性。合作者已同意测试用于研究的有用结构 在有生命的有机体中。 我们将研究该分子的复杂光动力学行为。这个 分子在吸收光谱中表现出可逆和不可逆的变化 这取决于辐射的波长。它还显示开/关开关 在适当的条件下和无氧条件下的行为， 当受到照射时，不可逆地变成红色荧光。的结构基础 这四种转变将在闪速冷冻晶体上通过 用于潜在生物和/或商业用途的结晶学技术。

Description: (Verbatim from the applicant's abstract) Biosensors designed to detect and provide a visible indication by changes in fluorescence of a variety of intracellular conditions will be developed by genetic engineering using the green fluorescent protein from the jellyfish Aequorea victoria as a template. Sensors have already been developed that respond to changes in pH and halide (chloride) concentration, which are essential for proper cellular function. These will be refined in function for use by cell and molecular biologists in single living cells, including human cells, in order to ease study of cellular processes in normal and diseased cells. In addition it is proposed to develop variants that are sensitive to changes in redox potential via reversible formation or breakage of disulfide bridges for use of detecting redox potentials in subcellular compartments such as the endoplasmic reticulum. Variants will be characterized by absorbance and fluorescence spectroscopy and the structures of potentially useful biosensors will be determined by X-ray crystallography in order to assess the structural basis for the molecular properties. Collaborators have agreed to test useful constructions for research in living organisms. The complex photodynamic behavior of the molecule will be investigated. The molecule shows reversible and irreversible changes in the absorption spectrum depending on the wavelength of irradiation. It also shows on/off switching behavior under appropriate conditions and in the absence of oxygen, irreversibly becomes red fluorescent when irradiated. The structural basis for these four transformations will be determined on flash-frozen crystals by crystallographic techniques for potential biological and/or commercial uses.