Development of a DNA microarray for authentication of ginseng drugs based on 18S rRNA gene sequence.
Development of a DNA microarray for authentication of ginseng drugs based on 18S rRNA gene sequence.
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DOI：
10.1021/jf0732814
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发表时间：
2008-05
影响因子：
6.1
通讯作者：
Shu Zhu;H. Fushimi;K. Komatsu
中科院分区：
文献类型：
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作者：
Shu Zhu;H. Fushimi;K. Komatsu
人参是五加科人参属植物人参的地下部分，是东方最重要的中草药。以前，13个人参分类群的核18 S rRNA基因的核苷酸序列进行了测定，因为是特定的多态性核苷酸用于识别每个物种。基于核苷酸差异，建立了人参属植物和药物的DNA微阵列（PNX array）。设计并合成了35种特异性寡核苷酸探针，其中33种探针对应种属特异性核苷酸替换，2种探针作为阳性对照和阴性对照。种特异性探针的长度为23-26 bp，其中替换核苷酸位于中间部分。通过校正荧光强度的变化来点样Trifecta探针以保证准确性。从人参属植物和药物及其衍生的保健食品中扩增的部分18 S rRNA基因序列作为靶点与PNX阵列杂交。在最佳条件下杂交后，可检测到人参属各物种的特异性荧光图谱，分析结果可显示为条形码，便于快速区分。PNX阵列的建立为人参属植物、人参药材及其保健食品的鉴别提供了客观、可靠的方法。
Ginseng drugs, derived from underground parts of Panax species (Araliaceae), are the most important group of herbal medicines in the Orient. Previously, the nucleotide sequences of the nuclear 18S rRNA gene of 13 Panax taxa were determined, as were the specific polymorphic nucleotides for identification of each species. On the basis of the nucleotide difference, a DNA microarray (PNX array) was developed for the identification of various Panax plants and drugs. Thirty-five kinds of specific oligonucleotide were designed and synthesized as probes spotting on a decorated glass slide, which included 33 probes corresponding to the species-specific nucleotide substitutions and 2 probes as positive and negative controls. The species-specific probes were of 23-26 bp in length, in which the substitution nucleotide was located at the central part. Triplicate probes were spotted to warrant accuracy by correcting variation of fluorescent intensity. Partial 18S rRNA gene sequences amplified from Panax plants and drugs as well as their derived health foods were fluorescently labeled as targets to hybridize to the PNX array. After hybridization under optimal condition, specific fluorescent patterns were detected for each Panax species, and the analyzed results could be indicated as barcode patterns for quick distinction. The developed PNX array provided an objective and reliable method for the authentication of Panax plants and drugs as well as their derived health foods.