摘要
随着分子生药学的发展,DNA分子标记在中药基原鉴定中的优势日益显著。为了弥补现有标记在中药质量监管中应用的局限性,该团队自主设计了一种新型分子标记,命名为DNA特征序列(DSS),该标记是仅出现在某个特定分类单元的DNA序列,长度40 bp,鉴定准确率高。该文旨在筛选并验证可准确鉴别《中国药典》一部中收录的植物类药材DSS标记,建立标准核酸序列研制的操作规程,为聚合酶链式反应法在中药质量监管中的广泛应用奠定基础。首先使用植物叶绿体基因组数据库(CGIR)获取待测样品、同属近缘物种及通用背景物种的叶绿体基因组序列,利用IdenDSS软件获得待测物种DSS序列组及其特异性引物对,通过DNA提取、PCR扩增、测序、序列比对进行验证,共获得203种药材的DSS序列。将上述序列上传至中药分子鉴定平台(www.herbsdna.com)标准数据库中,建立了用于大宗药材鉴定的标准DSS序列存储库,为中药质量监管提供重要的工具。在此基础上,形成DSS标准核酸序列研制操作规程,为进一步以更多元化的基因组序列为基础进行DSS筛选与应用奠定基础。
With the development of molecular pharmacognosy,the advantages of DNA molecular markers in the identification of original plants of Chinese medicinal materials are becoming increasingly significant.To compensate for the limitations of existing markers in the quality supervision of Chinese medicinal materials,our team has independently designed a new molecular marker named DNA signature sequence(DSS).This marker is a nucleotide sequence that only appears in a specific taxonomic unit,with a length of 40 bp and high identification accuracy.This article aims to screen and verify the DSS markers that can accurately identify the original plants of the medicinal materials included in the Volume One of the Chinese Pharmacopoeia,establish the operating procedure for developing standard nucleotide sequences,and lay a foundation for the widespread application of polymerase chain reaction in the quality supervision of traditional Chinese medicine.Firstly,the Chloroplast Genome Information Resource(CGIR)was searched for the chloroplast genome sequences of the test samples,species of the same genus,and common background species.IdenDSS was used to obtain the DSS tags and specific identification primers of the tested species.After DNA extraction,PCR amplification,sequencing,and sequence alignments,a total of 203 DSS markers of Chinese medicinal materials were obtained for validation.The above sequences were uploaded to the Traditional Chinese Medicine Molecular Identification Platform(www.herbsdna.com),and a standard DSS library was established for identifying the original plants of medicinal materials,serving as an important tool for quality supervision o f Chinese materia medica.On this basis,an operating procedure for DSS development is formed,laying a foundation for further DSS screening and application based on more diverse genome sequences.
作者
王多梅
蒋超
蒲婧哲
胡冲
陈灵丽
张亚中
袁媛
WANG Duo-mei;JIANG Chao;PU Jing-zhe;HU Chong;CHEN Ling-li;ZHANG Ya-zhong;YUAN Yuan(Anhui Institute of Food and Drug Control,Hefei 230051,China;NMPA Key Laboratory for Quality Research and Evaluation of Traditional Chinese Medicine,Hefei 230051,China;National Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第23期6249-6256,共8页
China Journal of Chinese Materia Medica
基金
安徽省药品监督管理局监管科学研究重点项目(AHYJ-KJ-202308)
安徽省药品监督管理局科技创新项目(AHYJ-KJ-202313)。
关键词
DNA特征序列
基原鉴定
标准核酸序列
DNA signature sequence
identification of original plants
standard nucleotide sequence
