Identification of Regulatory Protein Genes Involved in Alkaloid Biosynthesis Using a Transient RNAi System

Yasuyuki Yamada,Qingyun Luo,Joseph Gogo Dubouzet,Yasuhisa Kokabu,Nobuhiko Kato,Fumihiko Sato

doi:10.1007/978-1-60761-723-5_3

Identification of Regulatory Protein Genes Involved in Alkaloid Biosynthesis Using a Transient RNAi System

Yasuyuki Yamada, Qingyun Luo + Show 4 more

https://doi.org/10.1007/978-1-60761-723-5_3

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Publication Date: Jan 1, 2010

Citations: 2

Affiliation: Kyoto University, Nanjing Agricultural University, Scion

#Transient RNA Silencing #Candidate Regulatory Factors + Show 8 more

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Abstract

RNAi (RNA interference, RNA silencing) is a powerful tool in functional genomics. We report here the use of transient RNAi to isolate regulatory factor genes involved in isoquinoline alkaloid biosynthesis in Coptis japonica protoplasts. Double-stranded (ds) RNAs prepared against candidate regulatory factors, which were predicted from an EST library, were introduced into C. japonica protoplasts by polyethylene glycol (PEG)-mediated transformation, and their effects were monitored by real-time reverse transcription (RT)-polymerase chain reaction (PCR). The potential of this transient RNAi system to characterize the functions of regulatory factor genes in alkaloid research is discussed.

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