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719 bp,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99.4%,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。
Objective In order to clarify the phylogenetic relationship of one species of Uncaria sinensis（Oliv.） Havil,molecular identification and genetic analysis were carried out by 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 thereby rDNA ITS regions were amplified with universal primer,sequenced and phylogenetic analysed with Clustal X,BioEdit and PAUP.Results The entire rDNA ITS sequence of Uncaria was 719 bp.Sequence analysis showed that the rDNA ITS sequence was closely related to Uncaria sinensis（Oliv.） Havil available in GenBank and the similarity reached 99.4%.Conclusion Based on molecular biology methods of rDNA ITS region analysis,molecular identification is useful in accurate classification on Uncaria sinensis（Oliv.） Havil.,which provides molecular evidence of taxonomy and identification of different species in genus Uncaria.
出处
《广东药学院学报》
CAS
2012年第1期29-32,共4页
Academic Journal of Guangdong College of Pharmacy
基金
广东省自然科学基金(8451022401001607)
广东省中山市科技局资助项目(20101H017)
关键词
华钩藤
ITS序列
分子鉴定
Uncaria sinensis
ITS Sequence
molecular identification
