Species identification from Ginseng drugs by multiplex amplification refractory mutation system (MARMS).
Species identification from Ginseng drugs by multiplex amplification refractory mutation system (MARMS).
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DOI：
10.1055/s-2004-815502
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发表时间：
2004-02
期刊：
影响因子：
2.7
通讯作者：
Shu Zhu;H. Fushimi;Shaoqing Cai;K. Komatsu
中科院分区：
文献类型：
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作者：
Shu Zhu;H. Fushimi;Shaoqing Cai;K. Komatsu
采用多重扩增难突变系统（MARMS）对5种人参（参、参、西洋参、三七和越南参）进行了鉴定。根据叶绿体trnK基因和细胞核18S rRNA基因区域的特异性序列，为每个物种设计并合成了一套特异性引物，包括叶绿体trnK基因和细胞核18S rRNA基因区域的2对引物。利用5组特异性引物，依次以5个人参种提取的总DNA为模板，在适当条件下进行PCR扩增，每个产物进行琼脂糖凝胶电泳检测。结果表明，只有当一组物种特异性引物遇到相应物种的模板DNA时，才能同时观察到两个预期片段，一个来自trnK基因区域，另一个来自18S rRNA基因区域。该方法通过同时检测2个完全不同基因的4位核苷酸差异，不仅对5种人参品种的鉴定，而且对相应人参药物的鉴定结果更可靠。
The multiplex amplification refractory mutation system (MARMS) was applied to the identification of 5 Panax species ( P. ginseng, P. japonicus, P. quinquefolius, P. notoginseng and P. vietnamensis). A set of specific primers, including 2-pair primers on chloroplast trnK gene and nuclear 18S rRNA gene regions, respectively, was designed and synthesized for each species on the basis of species-specific sequences of the 2 genes. By using 5 sets of specific primers, in turn, PCR amplifications were performed with total DNA extracted from 5 Panax species as template under appropriate condition, and each resulting product was detected by agarose gel electrophoresis. The results showed that two expected fragments, one from trnK gene and another from 18S rRNA gene regions, were observed simultaneously only when the set of species-specific primers encountered template DNA of the corresponding species. This assay could give more reliable results for identification of not only 5 Panax species but also corresponding Ginseng drugs by simultaneous detection of 4-site nucleotide differences on 2 completely different genes.