目的常用生药柴胡正品为柴胡属植物柴胡及狭叶柴胡的干燥根。当前,同属多种植物也做柴胡内转录间隔区使用。仅凭生药性状及显微特征难以进行真伪鉴别。在本实验中,笔者针对6种不同柴胡属植物来源的柴胡生药,基于其内转录间隔区(ITS1)区多态性研制了用于其鉴别的DNA芯片。方法将设计的特异性寡核苷酸探针点印于基质上制备芯片;从待检测样品中提取基因组DNA,扩增ITS1片段,并以荧光标记后,与芯片杂交。结果在与其相应的探针位置即可出现信号;每种不同来源的柴胡有其特异的杂交谱图。结论研制的DNA芯片可为柴胡类生药的鉴定提供可靠便利的途径。
OBJECTIVE To develop an oligonucleotide DNA chip on the basis of the polymorphism of ITS1 region for identifying the 6 original species of Radix Bupleuri.METHODS Specie-specific oligonucleotide probes were printed onto substrate to fabricate a chip.The fluorescence-labeled ITS1 fragment amplified from genomic DNA of certain kind of Chaihu was then hybridized to the chip.RESULTS Each kind of Chaihu had its unique hybridization profile.Signals were obtained at places of its corresponding probes.CONCLUSION The chip can provide us with a credible and powerful tool for the identification of Chaihu.