SGER: Monitoring Cytoplasmic and Intraorganellar pH in Plants with a Novel BRET Reporter

SGER:利用新型 BRET 报告基因监测植物细胞质和细胞器内 pH 值

基本信息

  • 批准号:
    0854942
  • 负责人:
  • 金额:
    $ 15万
  • 依托单位:
  • 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:
    2009-02-15 至 2011-01-31
  • 项目状态:
    已结题

项目摘要

Regulation of intracellular acidity (pH) is vital to cells for many reasons. The measurement of pH inside living cells is therefore crucial to the study of many biological processes. The currently available methods for measuring intracellular pH mostly use fluorescence technologies that have limitations such as autofluorescence, phototoxicity, photobleaching, etc. These problems can be particularly serious in green plant tissue. Additionally, the currently available pH methods are useless when measuring light-dependent pH changes (such as those occurring in the chloroplast) because the method can trigger the very responses that are to be studied. The goals of this project is to develop a new method for measuring pH in living cells. The experimental approach that will be employed is based on a technique developed by the Principal Investigator of the project that allows for monitoring of in vivo protein interactions, Bioluminescence Resonance Energy Transfer (BRET). BRET avoids the problems of fluorescence methods. The project will develop a genetically encodable, targetable, ratiometric pH monitoring method. Therefore, pH can be quantified in living plant and animal cells with this method. The usefulness of this transformative technology will be demonstrated by application to important experimental questions in plant physiology, including the measurement of (i) pH changes in the chloroplast in response to light/dark transitions, and (ii) daily rhythms of cellular pH.Broader impacts.This project will lead to new information about the impact of the environment (for example, light/dark) on cellular activities in plants, thereby enabling new strategies for agricultural adaptation. The project will also provide an optimal training environment for a postdoctoral fellow and a graduate student to learn innovative technology development.
由于许多原因,细胞内酸度(pH)的调节对细胞至关重要。因此,活细胞内pH值的测量对于许多生物过程的研究至关重要。目前可用的用于测量细胞内pH的方法大多使用荧光技术,其具有诸如自发荧光、光毒性、光漂白等的局限性。这些问题在绿色植物组织中可能特别严重。此外,目前可用的pH方法在测量光依赖性pH变化(例如叶绿体中发生的pH变化)时是无用的,因为该方法可以触发要研究的响应。该项目的目标是开发一种测量活细胞pH值的新方法。将采用的实验方法是基于该项目的主要研究者开发的一种技术,该技术允许监测体内蛋白质相互作用,即生物发光共振能量转移(BRET)。BRET避免了荧光方法的问题。该项目将开发一种可遗传编码的、有针对性的比率pH监测方法。因此,可以用这种方法在活的植物和动物细胞中定量pH。这项变革性技术的有用性将通过应用于植物生理学中的重要实验问题来证明,包括测量(i)响应光/暗转换的叶绿体pH值变化,以及(ii)细胞pH值的每日节律。更广泛的影响。该项目将带来有关环境影响的新信息(例如,光/暗)对植物细胞活动的影响,从而使农业适应的新战略成为可能。该项目还将为博士后研究员和研究生学习创新技术开发提供最佳培训环境。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

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Carl Johnson其他文献

Videotape as a replacement for the human observer in studies of nonhuman primate behavior
在非人类灵长类动物行为研究中录像带替代人类观察者
  • DOI:
  • 发表时间:
    1972
  • 期刊:
  • 影响因子:
    0
  • 作者:
    D. K. Candland;L. Dresdale;J. Leiphart;Carl Johnson
  • 通讯作者:
    Carl Johnson
Cutaneous microthrombi: a histologic clue to the diagnosis of hypereosinophilic syndrome.
皮肤微血栓:诊断嗜酸性粒细胞增多综合征的组织学线索。
  • DOI:
  • 发表时间:
    1987
  • 期刊:
  • 影响因子:
    1.1
  • 作者:
    James E. Fitzpatrick;Carl Johnson;M. C. Major;Paul Simon;James Owenby;L. Colonel
  • 通讯作者:
    L. Colonel
10°C static storage mitigates ischemia reperfusion injury through improved mitochondrial preservation in a porcine DCD liver model
在猪 DCD 肝模型中,10°C 静态保存通过改善线粒体保存减轻缺血再灌注损伤
  • DOI:
    10.1016/j.ajt.2024.12.160
  • 发表时间:
    2025-01-01
  • 期刊:
  • 影响因子:
    8.200
  • 作者:
    Kaitlyn Tracy;Yutaka Shishido;Mark Petrovic;TiOluwanimi Adesanya;Avery Fortier;Timothy Harris;Michael Cortelli;William Tucker;Sean François;Brandon Petree;Kimya Raietparvar;Victoria Simon;Carl Johnson;Elizabeth Simonds;Nancy Cardwell;Christian Crannell;Alexandria Murphy;Melanie McReynolds;Ciara Shaver;Caitlin Demarest;Matthew Bacchetta
  • 通讯作者:
    Matthew Bacchetta
Towards the Development of a Pediatric Ventricular Assist Device
开发儿科心室辅助装置
  • DOI:
  • 发表时间:
    2006
  • 期刊:
  • 影响因子:
    3.3
  • 作者:
    H. Borovetz;S. Badylak;J. Boston;Carl Johnson;R. Kormos;M. Kameneva;M. Simaan;T. Snyder;H. Tsukui;W. Wagner;J. Woolley;J. Antaki;C. Diao;S. Vandenberghe;B. Keller;V. Morell;P. Wearden;S. Webber;J. Gardiner;Chung M. Li;Dave Paden;B. Paden;Shaun Snyder;Jingchun Wu;G. Bearnson;J. Hawkins;G. Jacobs;John Kirk;P. Khanwilkar;P. Kouretas;J. Long;R. Shaddy
  • 通讯作者:
    R. Shaddy
Voitenko experiments with novel diagnostics detect velocities of 89 km/s
  • DOI:
    10.1016/j.ijimpeng.2019.103406
  • 发表时间:
    2020-01-01
  • 期刊:
  • 影响因子:
  • 作者:
    Douglas G. Tasker;Young K. Bae;Carl Johnson;Kevin Rainey;Christopher Campbell;Dave Oschwald;Cornelius Reed
  • 通讯作者:
    Cornelius Reed

Carl Johnson的其他文献

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{{ truncateString('Carl Johnson', 18)}}的其他基金

Circadian Programming of Gene Expression in Cyanobacteria
蓝藻基因表达的昼夜节律编程
  • 批准号:
    9874371
  • 财政年份:
    1999
  • 资助金额:
    $ 15万
  • 项目类别:
    Continuing Grant
Development of Chemoenzymatic Routes to Bioactive Molecules
生物活性分子化学酶途径的开发
  • 批准号:
    9801679
  • 财政年份:
    1998
  • 资助金额:
    $ 15万
  • 项目类别:
    Continuing Grant
U.S.-Czech Research on Involvement of Melatonin and Calcium in Plant Photoperiodism
美国-捷克关于褪黑素和钙参与植物光周期的研究
  • 批准号:
    9605193
  • 财政年份:
    1997
  • 资助金额:
    $ 15万
  • 项目类别:
    Standard Grant
Circadian Rhythms of Gene Expression in Cyanobacteria
蓝藻基因表达的昼夜节律
  • 批准号:
    9633267
  • 财政年份:
    1996
  • 资助金额:
    $ 15万
  • 项目类别:
    Continuing Grant
U.S.-Japan Cooperative Research: Circadian Rhythms of Gene Expression in Cyanobacteria
美日合作研究:蓝藻基因表达的昼夜节律
  • 批准号:
    9218744
  • 财政年份:
    1993
  • 资助金额:
    $ 15万
  • 项目类别:
    Standard Grant
"Collaborative Research: Circadian Rhythms of Gene Expression in Cyanobacteria"
“合作研究:蓝藻基因表达的昼夜节律”
  • 批准号:
    9219880
  • 财政年份:
    1993
  • 资助金额:
    $ 15万
  • 项目类别:
    Continuing Grant
Development of Chemoenzymatic Approaches to Bioactive Molecules
生物活性分子化学酶方法的发展
  • 批准号:
    9223011
  • 财政年份:
    1993
  • 资助金额:
    $ 15万
  • 项目类别:
    Continuing Grant
Development of Chemoenzymatic Approaches to Bioactive Molecules
生物活性分子化学酶方法的发展
  • 批准号:
    8922955
  • 财政年份:
    1990
  • 资助金额:
    $ 15万
  • 项目类别:
    Continuing Grant
Synthetic Methodology Based on Sulfur and Organometallic Reagents (Chemistry)
基于硫和有机金属试剂的合成方法(化学)
  • 批准号:
    8607956
  • 财政年份:
    1986
  • 资助金额:
    $ 15万
  • 项目类别:
    Continuing Grant
Synthetic Methodology Based on Sulfur and Phosphorus Reagents (Chemistry)
基于硫磷试剂的合成方法(化学)
  • 批准号:
    8306594
  • 财政年份:
    1983
  • 资助金额:
    $ 15万
  • 项目类别:
    Continuing Grant

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