Chen Gu, Wang Xiao Li, Wong Wai Shing, Liu Xiao Dong, Xia Bing, Li Ning
Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong SAR, China.
J Agric Food Chem. 2005 Nov 2;53(22):8551-6. doi: 10.1021/jf051334g.
Radix astragali (root of Astragalus membranaceus) is an important traditional Chinese medicine. It has been used as a tonic herb for thousands of years in China. The water extract of the roots has a wide range of immunopotentiating effects and has been proven to be efficacious as an adjunct cancer therapy. Authentication of the herbal plant is routinely required for general practice in the field of herbal medicine. To facilitate rapid identification of numerous varieties of Radix astragali that are circulating on the herb markets, a rapid molecular genetic method, named 3' untranslated region (3' UTR) sequence-based amplified polymorphism (UAP), has been developed. A cDNA library was first built from transcripts of an authentic A. membranaceus species. Several cDNA clones specific to A. membranaceus were identified through subtractive hybridization of the A. membranaceus cDNA library with Arabidopsis total cellular RNA. On the basis of these cDNA sequences of the 3' untranslated region (3' UTR) of selected cDNA clones, a Polymerase Chain Reaction (PCR) was performed on genomic DNAs of the dry roots of several putative A. membranaceus. PCR fragment length polymorphism was found between A. membranaceus and its relatives. By using this method, it was possible to differentiate the authentic A. membranaceus root from those putative ones obtained from herbal medicine markets. To the authors' knowledge, this is the first paper applying UAP in the authentication of traditional Chinese medicine plants.
黄芪（膜荚黄芪的根）是一种重要的传统中药。在中国，它作为滋补草药已被使用了数千年。其根的水提取物具有广泛的免疫增强作用，并且已被证明作为辅助癌症治疗有效。草药领域的常规实践通常需要对草药植物进行鉴定。为了便于快速鉴定在草药市场上流通的众多黄芪品种，已经开发了一种快速分子遗传学方法，即基于3'非翻译区（3'UTR）序列的扩增多态性（UAP）。首先从正宗膜荚黄芪物种的转录本构建cDNA文库。通过膜荚黄芪cDNA文库与拟南芥总细胞RNA的消减杂交，鉴定出几个膜荚黄芪特有的cDNA克隆。基于所选cDNA克隆的3'非翻译区（3'UTR）的这些cDNA序列，对几种推定的膜荚黄芪干燥根的基因组DNA进行聚合酶链反应（PCR）。在膜荚黄芪与其近缘种之间发现了PCR片段长度多态性。通过使用这种方法，可以将正宗的膜荚黄芪根与从草药市场获得的那些推定的根区分开来。据作者所知，这是第一篇将UAP应用于中药植物鉴定的论文。
