Using STJs to measure the expression and interactions of Arabidopsis fluorescently-labelled proteins involved in pollen development

使用 STJ 测量参与花粉发育的拟南芥荧光标记蛋白的表达和相互作用

基本信息

  • 批准号:
    BB/E000258/1
  • 负责人:
  • 金额:
    $ 18.08万
  • 依托单位:
  • 依托单位国家:
    英国
  • 项目类别:
    Research Grant
  • 财政年份:
    2006
  • 资助国家:
    英国
  • 起止时间:
    2006 至 无数据
  • 项目状态:
    已结题

项目摘要

Pollen development is a critical step in the plant life cycle with commercial importance in crop breeding and seed production. We have been investigating the role that the MS1 protein, which is vital for pollen formation, plays in this process. We have identified a number of proteins that MS1 appears to interact with, but need to verify these interactions in planta. To do this we need to show when and where these proteins are expressed and confirm that they interact with MS1. One way to do this is to make fusion proteins combining a fluorescent marker to the protein of interest and to look for fluorescence and whether there is a change in the fluorescence as a consequence of interacting with the MS1 protein labelled with a different fluorescent marker. Detection of fluorescently labelled proteins are usually conducted by confocal microscopy, however, this is very difficult as the proteins are expressed in a limited number of cells within a tissue, chlorophyll autofluorescence partly masks the fusion protein fluorescence, and detecting small changes in fluorescence is extremely difficult. Although these issues apply to our work they are also common limitations encountered by other Plant Science researchers. One approach that may alleviate many of these problems is to use a novel form of detector (STJ) that detects the energy of individual photons. STJs for optical wavelengths have been developed by the European Space Agency and applied to the detection of biological fluorescence at the University of Leicester Space Research Centre. STJs measure the energies of individual optical photons with high efficiency and extremely low internal background. The detector offers an advantage over other photon counting systems because the entire fluorescent spectrum is detected on an event-by-event basis. This means that the contributions of different fluorescent labels can be distinguished without the need for filters and problems of autofluorescence can be eliminated. We wish to apply this technique to the detection and interaction of fluorescently labelled proteins in the anthers of a higher plant. This will validate this technique for the study of in vitro expression and interaction of proteins in plants. The use of STJ-based detection systems in plant biology may overcome many of the limitations that are currently encountered in the visualisation of fluorescent molecules in planta. This work will also provide valuable information on the process of pollen development.
花粉发育是植物生命周期中的关键步骤,在作物育种和种子生产中具有重要的商业意义。我们一直在调查的作用,MS1蛋白,这是至关重要的花粉形成,在这一过程中发挥作用。我们已经确定了一些蛋白质,MS1似乎相互作用,但需要验证这些相互作用在植物。要做到这一点,我们需要显示这些蛋白质在何时何地表达,并确认它们与MS1相互作用。这样做的一种方法是制备将荧光标记物与感兴趣的蛋白质结合的融合蛋白,并寻找荧光以及是否存在由于与用不同荧光标记物标记的MS 1蛋白质相互作用而导致的荧光变化。荧光标记的蛋白质的检测通常通过共聚焦显微镜进行,然而,这是非常困难的,因为蛋白质在组织内的有限数量的细胞中表达,叶绿素自发荧光部分掩盖了融合蛋白的荧光,并且检测荧光的微小变化是非常困难的。虽然这些问题适用于我们的工作,但它们也是其他植物科学研究人员遇到的常见限制。可以缓解这些问题中的许多问题的一种方法是使用检测单个光子的能量的新型检测器(STJ)。欧洲航天局开发了光波长的STJ,并将其应用于莱斯特大学空间研究中心的生物荧光检测。STJ以高效率和极低的内部背景测量单个光子的能量。该检测器提供了优于其他光子计数系统的优势,因为整个荧光光谱是在逐个事件的基础上检测的。这意味着可以区分不同荧光标记的贡献,而不需要过滤器,并且可以消除自发荧光的问题。我们希望将这一技术应用于高等植物花药中荧光标记蛋白的检测和相互作用。这将验证该技术在植物蛋白质体外表达和相互作用的研究。在植物生物学中使用基于STJ的检测系统可以克服目前在植物中荧光分子的可视化中遇到的许多限制。这项工作也将为花粉发育过程提供有价值的信息。

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)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Zoe Wilson其他文献

Zoe Wilson的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('Zoe Wilson', 18)}}的其他基金

Maintaining rice reproduction under high temperature stress:- identifying mechanisms and germplasm to increase crop resilience.
在高温胁迫下维持水稻繁殖:-确定提高作物恢复力的机制和种质。
  • 批准号:
    BB/N013700/1
  • 财政年份:
    2016
  • 资助金额:
    $ 18.08万
  • 项目类别:
    Research Grant
Developing systems to control male fertility in wheat for hybrid breeding, enhanced pollen production and increased yield.
开发控制小麦雄性育性的系统,用于杂交育种,提高花粉产量并提高产量。
  • 批准号:
    BB/P002080/1
  • 财政年份:
    2016
  • 资助金额:
    $ 18.08万
  • 项目类别:
    Research Grant
BBSRC Embrapa - Developing mitochondrial mediated resilience to abiotic stress during plant reproduction.
BBSRC Embrapa - 开发植物繁殖过程中线粒体介导的非生物胁迫恢复能力。
  • 批准号:
    BB/N004523/1
  • 财政年份:
    2015
  • 资助金额:
    $ 18.08万
  • 项目类别:
    Research Grant
Developing a Cereal Fertility Pipeline (CerFip) for wheat and barley.
开发小麦和大麦谷物肥力管道 (CerFip)。
  • 批准号:
    BB/J019666/1
  • 财政年份:
    2012
  • 资助金额:
    $ 18.08万
  • 项目类别:
    Research Grant
Systems analysis of tapetal regulatory networks required for pollen development
花粉发育所需绒毡层调控网络的系统分析
  • 批准号:
    BB/J001295/1
  • 财政年份:
    2011
  • 资助金额:
    $ 18.08万
  • 项目类别:
    Research Grant
Total Internal Reflection Fluorescence (TIRF) and Multidimensional Single Molecule Fluorescence (MSMM) microscopy of plant proteins
植物蛋白的全内反射荧光 (TIRF) 和多维单分子荧光 (MSMM) 显微镜
  • 批准号:
    BB/G000204/1
  • 财政年份:
    2009
  • 资助金额:
    $ 18.08万
  • 项目类别:
    Research Grant
Establishing an integrated network model for secondary wall thickening in anther development.
建立花药发育过程中次生壁增厚的综合网络模型。
  • 批准号:
    BB/F021062/1
  • 财政年份:
    2008
  • 资助金额:
    $ 18.08万
  • 项目类别:
    Research Grant
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了