Application of Single-Nucleotide Polymorphism Analysis of the trnK Gene to the Identification of Curcuma Plants
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概要
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We previously found that Curcuma plants and drugs derived from Curcuma longa, C. phaeocaulis, C. zedoaria, and C. aromatica could be identified by the nucleotide differences at two sites and the existence of a 4-base indel on trnK gene. In this paper, based on species-specific nucleotide sequences, the application of a new method, single-nucleotide polymorphism (SNP) analysis was investigated to identify Curcuma plants more conveniently. First, three types of reverse primer were synthesized in different lengths, 34 mer, 26 mer, and 30 mer, to anneal the template DNAs from each species at sites immediately upstream from substitution positions 177 and 645, and at the site including the 4-base insertion from 728 to 731, respectively. After single-base extension reaction of these primers using fluorescent-labeled ddNTPs and PCR products of the trnK gene region as template, the resulting products were detected using an ABI PRISM 310 Genetic Analyzer. The electrophoretogram showed three or two peaks at different positions depending on the 27 mer, 31 mer, and 35 mer product lengths. Each peak was derived from the incorporated fluorescent-labeled ddNMPs complementary to template nucleotides at positions 645, 724, and 177, respectively. C. phaeocaulis showed three peaks of ddCMP, ddAMP, and ddAMP. The other three species showed two peaks derived from 27 mer and 35 mer products: peaks of ddCMP and ddAMP in C. longa, those of ddCMP and ddTMP in C. zedoaria, and those of ddTMP and ddAMP in C. aromatica. Thus SNP analysis to identify four Curcuma plants was newly developed.
- 公益社団法人日本薬学会の論文
- 2004-01-01
著者
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SASAKI Yohei
Research Center for Ethnomedicines, Institute of Natural Medicine, Toyama Medical and Pharmaceutical
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FUSHIMI Hirotoshi
Research Institute for Wakan-Yaku (Traditional Sino-Japanese Medicines), Toyama Medical and Pharmace
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Sasaki Y
Tohoku Pharmaceutical University
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Sasaki Y
Institute Of Natural Medicine University Of Toyama
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Komatsu Katsuko
富山大学和漢医薬学総合研究所
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Komatsu K
Div. Of Pharmacognosy Dep. Of Medicinal Resources Inst. Of Natural Medicine Univ. Of Toyama
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Komatsu K
Institute Of Natural Medical Toyama Medicine And Pharmaceutical University
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Komatsu Katsuko
Division Of Pharmacognosy Department Of Medicinal Resources Institute Of Natural Medicine Toyama Med
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Komatsu Katsuko
Research Institute For Wakan-yaku (traditional Sino-japanese Medicines) Toyama Medical And Pharmaceu
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Komatsu Katsuko
Division Of Pharmacognosy Institute Of Natural Medicine University Of Toyama
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KOMATSU Katsuko
21st Century COE Program, Toyama Medical and Pharmaceutical University
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Komatsu K
Institute Of Natural Medicine University Of Toyama
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Fushimi Hirotoshi
Institute Of Natural Medicine University Of Toyama
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Fushimi Hirotoshi
Div. Of Pharmacognosy Dep. Of Medicinal Resources Inst. Of Natural Medicine Univ. Of Toyama
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Fushimi Hirotoshi
Research Institute For Wakan-yaku (traditional Sino-japanese Medicines) Toyama Medical And Pharmaceu
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Sasaki Yohei
Research Center For Ethnomedicines Institute Of Natural Medicine Toyama Medical And Pharmaceutical U
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Sasaki Youhei
Faculty Of Pharmaceutical Sciences Hoshi University
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Sasaki Yohei
Department Of Medicinal Plant Science Hoshi University
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Fushimi Hirotoshi
Research Center for Ethnomedicines, Institute of Natural Medicine, Toyama Medical and Pharmaceutical University
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