DEVELOPMENT OF METABOLIC ENGINEERING OF PLANT ; CHLOROPLAST TECHNOLOGY OF CHILAMYDOMONAS

植物代谢工程的发展;

基本信息

  • 批准号:
    09450305
  • 负责人:
  • 金额:
    $ 8.32万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
  • 财政年份:
    1997
  • 资助国家:
    日本
  • 起止时间:
    1997 至 1999
  • 项目状态:
    已结题

项目摘要

The chloroplast is one of the most important organelles in plant cells. Genetic manipulation of the chloroplast genome might be an important technique for construction of useful transgenic plants in the future. In order to express a foreign gene efficiently in the chloroplast, we have to obtain a thorough understanding of the regulation of chloroplast gene expression.Chimeric genes for expression of a foreign gene in the Chlamydomonas reinhardtii chloroplast were constructed. There chimeric genes are composed of the promoter from chloroplast genes, rbcL, psbA, psbD, and atpA, 5'- and 3'-untranslated regions, and the Escherichia coli b-glucuronidase (GUS) structural gene idA) as a foreign gene. Four types of chloroplast transformants (RG, PG, PDG, and AG) were generated. The AG transformant showed the highest GUS activity so far reported in C.reinhardtii, and the accumulated GUS protein accounted for 1.4% of the total soluble proteins. However, the pattern of gene expression was not the same as that of the endogenous genes in the chloroplast, in order to express a foreign gene efficiently in the chloroplast, it was necessary to fuse a coding region from chloroplast gene.We analyzed the effect of nucleoties immediately upstream of the initiation codon (AUG) on the translation efficiency. In the case of chmeric-uidA (atpA), the substitution of two nucleoties immediately upstream of the AUG changed the translation efficiency. On the other hand, in the case of chimeric-uidA (psbD), that had no effect. We also tried to construct a translation system from polycistronic mRNA, and partially successed.Study on the control mechanisms of gene expression of the Chlamydomonas chloroplast genome in detail will be applied to transformation of the chloroplast genome in higher agriculturally and industrially important plants in the near future.
叶绿体是植物细胞中最重要的细胞器之一。叶绿体基因组的遗传操作可能是构建有用转基因植物的重要技术。为了使外源基因在叶绿体中高效表达,我们必须深入了解叶绿体基因表达的调控机制。这些嵌合基因由来自叶绿体基因的启动子、rbcL、psbA、psbD和atpA、5 ′-和3 ′-非翻译区以及作为外源基因的大肠杆菌β-葡萄糖醛酸酶(GUS)结构基因idA组成。产生四种类型的叶绿体转化体(RG、PG、PDG和AG)。其中,AG基因的GUS活性最高,积累的GUS蛋白占总可溶性蛋白的1.4%。然而,外源基因在叶绿体中的表达模式与内源基因的表达模式不同,为了使外源基因在叶绿体中高效表达,需要融合叶绿体基因的编码区,本文分析了起始密码子(AUG)上游核苷酸对外源基因翻译效率的影响。在chmeric-uidA(atpA)的情况下,紧接着AUG上游的两个核苷酸的取代改变了翻译效率。另一方面,在嵌合uidA(psbD)的情况下,这没有影响。我们还尝试构建了多顺反子mRNA的翻译系统,并取得了部分成功。对衣原体叶绿体基因组表达调控机制的详细研究,将在不久的将来应用于高等农业和工业重要植物叶绿体基因组的转化。

项目成果

期刊论文数量(7)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Kiyohide Ishikura: "The coding regions of RuBisCO large subunit gene effects on its gene expression." Abstracts ; 5th International Congress of Plant Molecular Biology. 1190-1190 (1997)
Kiyohide Ishikura:“RuBisCO 大亚基基因的编码区对其基因表达有影响。”
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    0
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  • 通讯作者:
石倉清秀: "Expression of a Foreign Gene in Chlamydomonas reinhardfii Chloroplast" J.Biosci.Bioeng.87・3. (1999)
Kiyohide Ishikura:“莱茵衣藻叶绿体中外源基因的表达”J.Biosci.Bioeng.87・3(1999)。
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    0
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Ko Kato: "Functional expression of a foreign eukaryotic ribulose-1,5-bisphosphate carboxylase/oxygerase large subunit gene in Chlamydomonas chloroplast" Abstracts ; 5th International Congress of Plant Molecular Biology. 491-491 (1997)
Ko Kato:“外源真核核酮糖-1,5-二磷酸羧化酶/加氧酶大亚基基因在衣藻叶绿体中的功能表达”摘要;
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  • 影响因子:
    0
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  • 通讯作者:
Ishikura Kiyohide: "Expression of a Foreign Gene in Chlamydomonas reinhardtii chloroplast"Journal of Bioscience and Bioengineering. 87・3. 307-314 (1999)
石仓清英:“莱茵衣藻叶绿体中的外源基因的表达”《生物科学与生物工程杂志》87·3(1999)。
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    0
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SHINMYO Atsuhiko其他文献

SHINMYO Atsuhiko的其他文献

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{{ truncateString('SHINMYO Atsuhiko', 18)}}的其他基金

Application of Salt-tolerant and Nitrogen-fixing Blue-green Algae to Agriculture in Sea Water
耐盐固氮蓝绿藻在海水农业中的应用
  • 批准号:
    14205118
  • 财政年份:
    2002
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
System for Screening for microorganisms with specific functions or specific microbial products
特定功能微生物或特定微生物产品筛选系统
  • 批准号:
    11695050
  • 财政年份:
    1999
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Application of no utility plant resources by a recombinant DNA technology
重组DNA技术对无用植物资源的应用
  • 批准号:
    10555289
  • 财政年份:
    1998
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
METABOLIC ENGINEERING OF CULTURED TOBACCO CELLS
培养烟草细胞的代谢工程
  • 批准号:
    07555258
  • 财政年份:
    1995
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
CONSTRUCTION OF PRODUCTION SYSTEMS OF USEFUL COMPOUNDS IN TOBACCO CULTURED CELLS BY RECOMBINANT DNA TECHNOLOGY
利用重组DNA技术构建烟草培养细胞中有用化合物的生产体系
  • 批准号:
    05555221
  • 财政年份:
    1993
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Developmental Scientific Research (B)
CONSTRUCTION OF PRODUCTION SYSTEMS OF USEFUL COMPOUNDS IN PLANT CELL CULTURE BY APPLICATION OF PLANT GENE EXPRESSION SYSTEM
应用植物基因表达系统构建植物细胞培养有用化合物生产体系
  • 批准号:
    04454037
  • 财政年份:
    1992
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
Expression of Cucumber Ascorbate Oxidase gene in Microbes
黄瓜抗坏血酸氧化酶基因在微生物中的表达
  • 批准号:
    02650712
  • 财政年份:
    1990
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)

相似海外基金

Gene-drive system for efficient chloroplast transformation
用于高效叶绿体转化的基因驱动系统
  • 批准号:
    BB/R021937/1
  • 财政年份:
    2019
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Research Grant
Production of stress-tolerant wheat by chloroplast transformation
通过叶绿体转化生产抗逆小麦
  • 批准号:
    24658010
  • 财政年份:
    2012
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Genetic modification of Agrobacterium tumefaciens in order to use chloroplast transformation.
对根癌农杆菌进行遗传改造,以利用叶绿体转化。
  • 批准号:
    22658025
  • 财政年份:
    2010
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Functional analysis of plastid transcription factors by chloroplast transformation.
通过叶绿体转化对质体转录因子进行功能分析。
  • 批准号:
    14540598
  • 财政年份:
    2002
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Functional analysis of chloroplast genes by chloroplast transformation
通过叶绿体转化进行叶绿体基因的功能分析
  • 批准号:
    10440238
  • 财政年份:
    1998
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
Analysis of transcriptional regulation in chloroplasts by DNA-associated transcrition factors by means of chloroplast transformation
通过叶绿体转化分析 DNA 相关转录因子对叶绿体的转录调控
  • 批准号:
    10640629
  • 财政年份:
    1998
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Basic studies on the chloroplast transformation using photoautotrophic cultured cells
光合自养培养细胞叶绿体转化的基础研究
  • 批准号:
    63560082
  • 财政年份:
    1988
  • 资助金额:
    $ 8.32万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
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