CORE--MOTOR PROTEIN PRODUCTION

核心——运动蛋白的产生

基本信息

  • 批准号:
    6100628
  • 负责人:
  • 金额:
    $ 20.03万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    1999
  • 资助国家:
    美国
  • 起止时间:
    1999-07-15 至 2000-06-30
  • 项目状态:
    已结题

项目摘要

The function of Core B is to provide proteins and custom synthesized organic chemicals t the various projects. During the past four years the core has produced and distributed a number of proteins. These were produced by two technicians, Aaron Ruby, who recently left for graduate school, and Marija Matuska. Both kinesin and NCD monomers were produced and provided to Cooke, Mendelson Fletterick and Highsmith. NCD dimers were produced and distributed to Fletterick, Cooke, and Mendelson. Actin and myosin subfragments were distributed to Spudich and Highsmith. Aaron Ruby participated in the production of a series of kinesin mutants that were used in the Vale laboratory and will also be available for the other projects. Microtubules were produced and distributed to Cooke, Highsmith, Vale and Mendelson. The central production of these proteins has greatly accelerated the progress made by the program. The production of these proteins by each laboratory would be a time consuming process, that would waste time and supplies. The core also produced a fluorescent analog of GTP for use in the Vale and Cooke laboratories. The proteins to be produced fall into two categories, native proteins made from animal tissues, and proteins cloned and expressed in E. coli., Dictyostelium discoidium SF-9 or pichia systems. The first group includes kinesins, actin and microtubules and myosin. These protein purification follow established procedures, and usually the proteins will be used by two or more of the projects. Where feasible, proteins will be cloned and expressed in E. coli. Typically, due to favorable growth and expression characteristics, both wild type and mutant proteins will be used in the projects. Expression and purification of both wild type and mutant myosin subfragments will be from dictyostelium, and clones for this proteins will be constructed in the Spudich project. Once the expression of a given protein in the dictyostelium has been established, this strain of dictyostelium will be produced by personnel from the Core operating in the Spudich lab. Constructs for other proteins will derived from the Vale or Fletterick labs.
核心B的功能是为各种项目提供蛋白质和定制合成的有机化学品。在过去的四年里,核心已经产生和分布了许多蛋白质。这些是由两名技术人员制作的,他们是最近去读研究生的亚伦·鲁比和玛丽亚·马图斯卡。产生驱动蛋白和NCD单体并提供给Cooke、Mendelson Fletterick和Highsmith。生产NCD二聚体并分发给Fletterick、Cooke和Mendelson。肌动蛋白和肌球蛋白亚片段被分配给Spudich和Highsmith。Aaron Ruby参与了一系列驱动蛋白突变体的生产,这些突变体在Vale实验室中使用,也将用于其他项目。微管产生并分配给库克,海史密斯,淡水河谷和门德尔松。这些蛋白质的集中生产大大加快了该计划的进展。每个实验室生产这些蛋白质将是一个耗时的过程,这将浪费时间和物资。该核心还产生了一种GTP的荧光类似物,供Vale和Cooke实验室使用。待生产的蛋白质分为两类,从动物组织中制备的天然蛋白质和在大肠杆菌中克隆和表达的蛋白质。大肠杆菌,盘基网柄藻SF-9或毕赤酵母系统。第一组包括驱动蛋白、肌动蛋白、微管和肌球蛋白。这些蛋白质纯化遵循既定程序,通常蛋白质将用于两个或更多个项目。在可行的情况下,将在E.杆菌通常,由于有利的生长和表达特性,野生型和突变型蛋白质都将用于项目中。野生型和突变型肌球蛋白亚片段的表达和纯化将来自网骨藻,并且该蛋白的克隆将在Spudich项目中构建。一旦在网骨藻中建立了给定蛋白质的表达,这种网骨藻菌株将由在Spudich实验室工作的核心人员生产。其他蛋白质的构建体将来自Vale或Fletterick实验室。

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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Roger A Cooke其他文献

Roger A Cooke的其他文献

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{{ truncateString('Roger A Cooke', 18)}}的其他基金

Conformational Changes in Molecular Motors
分子马达的构象变化
  • 批准号:
    7468032
  • 财政年份:
    2007
  • 资助金额:
    $ 20.03万
  • 项目类别:
PURCHASE OF A BRUKER EMX EPR SPECTROMETER: PROTEINS
购买 BRUKER EMX EPR 光谱仪:蛋白质
  • 批准号:
    6973345
  • 财政年份:
    2004
  • 资助金额:
    $ 20.03万
  • 项目类别:
Conformational Changes in Molecular Motors-Roger Cooke
分子马达的构象变化-罗杰库克
  • 批准号:
    6837556
  • 财政年份:
    2004
  • 资助金额:
    $ 20.03万
  • 项目类别:
Purchase of a Bruker EMX EPR spectrometer
购买布鲁克 EMX EPR 光谱仪
  • 批准号:
    6731931
  • 财政年份:
    2004
  • 资助金额:
    $ 20.03万
  • 项目类别:
PURCHASE OF A BRUKER EMX EPR SPECTROMETER: YEAST PRION
购买 BRUKER EMX EPR 光谱仪:酵母朊病毒
  • 批准号:
    6973346
  • 财政年份:
    2004
  • 资助金额:
    $ 20.03万
  • 项目类别:
CONFORMATIONAL CHANGES IN MOLECULAR MOTORS
分子马达的构象变化
  • 批准号:
    6456691
  • 财政年份:
    2001
  • 资助金额:
    $ 20.03万
  • 项目类别:
CONFORMATIONAL CHANGES IN MOLECULAR MOTORS
分子马达的构象变化
  • 批准号:
    6347853
  • 财政年份:
    2000
  • 资助金额:
    $ 20.03万
  • 项目类别:
CONFORMATIONAL CHANGES IN MOLECULAR MOTORS
分子马达的构象变化
  • 批准号:
    6338665
  • 财政年份:
    2000
  • 资助金额:
    $ 20.03万
  • 项目类别:
CORE--MOTOR PROTEIN PRODUCTION
核心——运动蛋白的产生
  • 批准号:
    6338664
  • 财政年份:
    2000
  • 资助金额:
    $ 20.03万
  • 项目类别:
CONFORMATIONAL CHANGES IN MOLECULAR MOTORS USING PARAMAGNETIC PROBES
使用顺磁探针观察分子马达的构象变化
  • 批准号:
    6308886
  • 财政年份:
    2000
  • 资助金额:
    $ 20.03万
  • 项目类别:

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