Transient RNA Silencing of Scoulerine 9-O-Methyltransferase Expression by Double Stranded RNA in Coptis japonica Protoplasts
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概要
- 論文の詳細を見る
RNAi (RNA interference, RNA silencing) is a powerful tool for functional genomics, but the construction of an RNAi vector(s) and the establishment of stable transformants are time-consuming and laborious. Here we report the transient RNAi of endogenous biosynthetic genes involved in isoquinoline alkaloid biosynthesis in Coptis japonica protoplasts. Double stranded (ds) RNA fragments of various lengths prepared from several different positions of the coding sequence of scoulerine 9-O-methyltransferase (SMT) were introduced into C. japonica protoplasts by polyethylene glycol-mediated transformation, and their effects were monitored by reverse transcription-polymerase chain reaction. Substantial silencing of SMT gene expression was obtained by the introduction of these SMT dsRNAs. A significant reduction in SMT protein levels was also observed. The potentials of this transient RNAi system to evaluate the functions of biosynthetic genes in Coptis alkaloid research are discussed.
- 社団法人 日本農芸化学会の論文
- 2005-01-23
著者
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An Chung-il
Department Of Biotechnology Graduate School Of Engineering Osaka University
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FUKUSAKI EI-ICHIRO
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Sato Fumihiko
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
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IFUKU Kentaro
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University
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Sato Fumihiko
Div. Integ. Life Sci. Grad. Sch. Biostudies Kyoto Univ.:plant Function Lab.
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DUBOUZET Joseph
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University
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MORISHIGE Takashi
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University
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FUJII Nanae
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University
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Fukusaki Ei-ichiro
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Sato Fumihiko
Laboratory Of Molecular And Cellular Biology Of Totipotency Division Of Integrated Life Science Grad
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Fujii Nanae
Laboratory Of Molecular And Cellular Biology Of Totipotency Division Of Integrated Life Science Grad
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Ifuku Kentaro
Division Of Integrated Life Science Graduate School Of Biostudies Kyoto University
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Sato F
Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University
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Fujii N
Research And Development Division Kikkoman Corporation
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Dubouzet Joseph
Division Of Integrated Life Science Graduate School Of Biostudies Kyoto University
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Morishige Takashi
Laboratory Of Molecular And Cellular Biology Of Totipotency Division Of Integrated Life Science Grad
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Sato Fumihiko
Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University
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