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題 名 | Molecular Differentiation of Atractylodes Drugs by PCR-Restriction Fragment Length Polymorphism and PCR-Selective Restriction Analysis on the 18S-5.8S rDNA Intratranscribed Spacer 1 Gene=應用PCR-RFLP與PCR-SR方法分析朮屬中藥材18S-5.8S核醣體核酸基因之內轉錄第一間隔區作為該等藥材基原之鑑別 |
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作 者 | 陳惠芳; 賴本; 詹秀真; 周清邦; 楊德動; 黃文鴻; 廖俊亨; 林嘉伯; | 書刊名 | 藥物食品分析 |
卷 期 | 5:4 1997.12[民86.12] |
頁 次 | 頁319-327 |
專 輯 | 中藥專輯 |
分類號 | 414.32 |
關鍵詞 | 關蒼朮; 茅蒼朮; 白朮; 基原鑑定; 18S-5.8S核醣體核酸基因之內轉錄第一間隔區; Atractylodes japonica; Atractylodes lancea; Atractylodes ovata; ITS[feb5]; PCR-RFLP; PCR-SR; Herbal origin; |
語 文 | 英文(English) |
中文摘要 | 本研究目的乃在建立以分子生物技術鑑別朮屬(Atractylode)中藥材等之基原。藉由 聚合酵素鏈鎖反應方法(PCR)將關蒼朮(Atractylodes japonica)、茅蒼朮(Atractyldes lancea)、白 朮(Atractylodes ovata)等之18S-5.8S核醣體核酸基因之內轉錄第一間隔區(簡稱ITS )複製出, 由此三種藥材所複製出之PCR產物,其大小均約為320bp,以限制酵素MspI水解作用後, 於不同種藥材卻有不同大小之水解產物;進一步選殖其各自之PCR產物並進行核酸序列分 析。發現此三種藥材之ITS 核酸序列彼此均異,且有特殊之限制酵素選擇作用處可作彼此 之區分,如:只有茅蒼朮之ITS ,可被BspEI作用,而其它二種藥材之ITS ,其BspEI 作用處;反之,只有關蒼朮與白朮之ITS ,具有Af/II作用處,而茅蒼朮之ITS ,無該酵 素作用處;另外,僅白朮之ITS 具有NaeI作用處,而其它二種藥材之ITS 無NaeI用處。 因此,由本研究結果,發現利用PCR-RFLP與PCR-SR分析朮屬藥材之ITS 區域,可作為 該等藥材基原之鑑別。 |
英文摘要 | The 18S-5.8S rDNA intratranscribed spacer 1 (ITS ) regions from Atractylodes japonica, A. lancea and A. ovata rhizomes were amplified using the polymerase chain reaction (PCR) with consensus rRNA gene primers. The amplified DNA fragment from Atractylodes species were of similar size, but with different MspI restriction mapping. Cloning and sequencing of the PCR products showed that the DNA sequences of PCR-amplified ITS were distinct between Atractylodes species. Sequene analysis indicated that the amplified ITS elements were informative for ITS fingerprinting following digestion with 6-cutter restriction endonu-cleases BspEI, AflII, and NaeI. Therefore, both polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and polymerase chain reaction-selective restriction (PCR-SR) analysis on the ITS gene were established to provide useful tools for the authentication between Atractylodes rhizomes. |
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