Ribozymes for new genetic coding systems

用于新遗传编码系统的核酶

基本信息

  • 批准号:
    6844940
  • 负责人:
  • 金额:
    $ 32.85万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2000
  • 资助国家:
    美国
  • 起止时间:
    2000-02-01 至 2008-01-31
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this research is to devise a novel and practical tRNA aminoacylation system based on ribozymes, which facilitates the technique of nonnatural amino acid mutagenesis and thus raises it to a more user-accessible technology. An artificial ribozyme selected in our laboratory exhibits a broad spectrum of activities toward phenylalanine (Phe) analogs and suppressor tRNAs bearing different nonsense codons. By immobilizing this ribozyme (called PheFlexizyme) on a resin, the synthesis of suppressor tRNAs charged with Phe analogs is largely facilitated; where a user-specified tRNA and a Phe analog are reacted on this resin and the resulting filtrate (or supernatant) contains the desired aminoacyl-tRNA. This charged tRNA is then used in a cell-free translation system to incorporate a Phe analog at a single position or two Phe analogs at two specific positions. The entire processes, including tRNA charging, in vitro translation, and purification of protein, can be done in one day. The translation efficiency is generally 50-70 mu/g/mL, thus it is feasible to produce a sufficient amount of protein for further biological studies. It should be noted that the PheFlexizyme-resin can be reused more than 10 times, and therefore it is also economical. In this application, we will attempt to develop more sophisticated ribozyme aminoacylation technologies. Specific aims are (1) expanding repertoire of Phe analogs for PheFlexizyme, (2) in vitro evolution of new Flexizymes based on the PheFIlexizyme scaffold, (3) in situ generation of PheFlexizyme in the cell-free transcription-coupled translation apparatus, and (4) applications of PheFlexizyme.
描述(由申请人提供):

项目成果

期刊论文数量(0)
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John P Richard其他文献

John P Richard的其他文献

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{{ truncateString('John P Richard', 18)}}的其他基金

Studies on Enzyme Activation and Novel Modes of Inhibition
酶激活和新抑制模式的研究
  • 批准号:
    10317064
  • 财政年份:
    2020
  • 资助金额:
    $ 32.85万
  • 项目类别:
Studies on Enzyme Activation and Novel Modes of Inhibition
酶激活和新抑制模式的研究
  • 批准号:
    10543563
  • 财政年份:
    2020
  • 资助金额:
    $ 32.85万
  • 项目类别:
Activation of Enzymes for Catalysis: The Role of Substrate-Induced Structural Changes
催化酶的激活:底物诱导的结构变化的作用
  • 批准号:
    9198549
  • 财政年份:
    2016
  • 资助金额:
    $ 32.85万
  • 项目类别:
Ribozymes for new genetic coding systems
用于新遗传编码系统的核酶
  • 批准号:
    7188027
  • 财政年份:
    2000
  • 资助金额:
    $ 32.85万
  • 项目类别:
Ribozymes for new genetic coding systems
用于新遗传编码系统的核酶
  • 批准号:
    7012204
  • 财政年份:
    2000
  • 资助金额:
    $ 32.85万
  • 项目类别:
MECHANISMS FOR ENZYME CATALYSIS OF HETEROLYTIC REACTIONS
酶催化杂解反应的机制
  • 批准号:
    3306773
  • 财政年份:
    1992
  • 资助金额:
    $ 32.85万
  • 项目类别:
MECHANISMS FOR ENZYME CATALYSIS OF HETEROLYTIC REACTIONS
酶催化杂解反应的机制
  • 批准号:
    3306774
  • 财政年份:
    1992
  • 资助金额:
    $ 32.85万
  • 项目类别:
MECHANISMS FOR ENZYME CATALYSIS OF HETEROLYTIC REACTION
酶催化杂解反应的机理
  • 批准号:
    3306775
  • 财政年份:
    1992
  • 资助金额:
    $ 32.85万
  • 项目类别:
MECHANISMS FOR ENZYME CATALYSIS OF HETEROLYTIC REACTION
酶催化杂解反应的机理
  • 批准号:
    2184726
  • 财政年份:
    1992
  • 资助金额:
    $ 32.85万
  • 项目类别:
MECHANISMS FOR ENZYME CATALYSIS OF HETEROLYTIC REACTION
酶催化杂解反应的机理
  • 批准号:
    2184725
  • 财政年份:
    1992
  • 资助金额:
    $ 32.85万
  • 项目类别:
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