CAJ | 학술논문

为了确保该药材的质量及临床安全用药，需要对常用中药当归及其混伪品进行分子鉴定。本文介绍了利用 ITS2条形码进行鉴定的方法：对 ITS2序列进行 PCR 扩增，并将纯化后的 PCR 产物进行双向测序，运用CodonCode Aligner拼接后，基于HMMer注释方法去掉两端的5.8S区和28S区获得ITS2条形码，用MEGA5.0软件进行相关数据分析，构建NJ树。结果表明：当归与其混伪品ITS2条形码间存在明显差异，基于ITS2条形码构建的NJ树能够将当归及其混伪品区分开。因此， ITS2条形码可有效地鉴别中药当归及其混伪品，为保证其临床用药安全提供了新的技术手段。
위료학보해약재적질량급림상안전용약，수요대상용중약당귀급기혼위품진행분자감정。본문개소료이용 ITS2조형마진행감정적방법：대 ITS2서렬진행 PCR 확증，병장순화후적 PCR 산물진행쌍향측서，운용CodonCode Aligner병접후，기우HMMer주석방법거도량단적5.8S구화28S구획득ITS2조형마，용MEGA5.0연건진행상관수거분석，구건NJ수。결과표명：당귀여기혼위품ITS2조형마간존재명현차이，기우ITS2조형마구건적NJ수능구장당귀급기혼위품구분개。인차， ITS2조형마가유효지감별중약당귀급기혼위품，위보증기림상용약안전제공료신적기술수단。
In this paper, a method using ITS2 barcode to identify Angelicae sinensis Radix and its adulterants is introduced. Firstly, the PCR technique was performed to get the ITS2 barcodes and puriifed PCR products were sequenced bi-directionally. Then sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. To remove the 5.8S and 28S region, the hidden Markov model (HMM)-based annotation methods were used. Finally, the ITS2 barcodes were aligned with Clustal-W and the genetic distances were computed by MEGA 5.0 according to the Kimura-2-Parameter (K2P) model. The Neighbor-joining (NJ) phylogenetic tree was constructed. The results showed that there is obvious genetic divergence in the ITS2 sequences between Angelicae sinensis Radix and its adulterants. NJ tree based on ITS2 barcodes can distinguish Angelicae sinensis Radix and its adulterants. This concludes that ITS2 region as a DNA barcode can effectively provided a new technique for identiifcation of Angelicae sinensis Radix.