一种贵州华钩藤的分子鉴定.docVIP

精品论文 参考文献 一种贵州华钩藤的分子鉴定 周子雄1 周林2 杜贵锋2 冯兵芳2 谢君婷2 朱爽(通讯作者)2 　　（1广东药学院中药学院 广东 广州 510006） 　　（2广东药学院生命科学与生物制药学院 广东省生物技术候选药物研究重点实验室 广东 广州 510006） 　　【摘要】 目的：以核糖体转录间隔区(rDNA ITS)序列为分子标记，对1种产自贵州安顺的华钩藤植物进行遗传分析与分子鉴定。方法：通过改良CTAB法提取样品总DNA，利用通用引物对rDNA ITS序列进行PCR扩增，经克隆、测序后，运用Clustal X、BioEdit和PAUP等软件进行序列分析和构建系统发育树。结果：测序得到样品的rDNA ITS区序列长度为719bp，序列分析结果显示其与Genbank中已有的华钩藤[Uncaria sinensi(Oliv.)Havil.] rDNA ITS区序列之间相似性达99.8%，并且在系统发育树中并排聚类成1支。结论：基于rDNA ITS区序列的测序分析和系统发育树构建的分子生物学方法，能够对华钩藤植物进行准确的分子鉴定，为钩藤属中药材的种类鉴定和种间分类提供分子生物学理论依据。 　　【关键词】 华钩藤；ITS序列；分子鉴定 　　【中图分类号】R927.2 【文献标识码】A 【文章编号】2095-1752（2015）08-0098-03 　　Molecular identification of one Guizhou 　　Uncaria sinensis(Oliv.)Havil. 　　Zhou Zixiong1, Du Guifeng2, Feng Bingfang2 XIie Gunting2, Zhu Shuang2* 　　(1.School of Traditional Chinese Medicine, Guangdong Pharmaceutical University, Guangzhou510006,China；2. Guangdong Province Key Laboratory for Biotechnology Drug Candidates, School of Biosciences and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou510006,China) 　　【Abstract】Objective:To characterize the phylogenetic relationship of one Guizhou Uncaria sinensis(Oliv.)Havil,molecular identification and genetic analysis were carried out using the rDNA ITS sequence as molecular marker. Methods:Molecular identification and genetic analysis were carried out with the rDNA ITS sequence as molecular marker.Total DNA was extracted with modified CTAB Method:and there by rDNA ITS regions were amplified with primers ITS4/ITS5, sequenced and phylogenetic analysed with Clustal X, BioEdit and PAUP. Results:The entire rDNA ITS sequence of Uncaria was 719bp.Sequence analysis showed that the rDNA ITS sequence was closely related to Uncaria sinensis(Oliv. ) Havil available in GenBank and the similarity reached 99.8%. Conclusion Based on molecular biology Methods:of rDNA ITS region analysis, molecular identification is useful inaccurate classification on Uncaria sinensis(Oliv.)Havil.,which provides m