基于COⅠ与16S rRNA基因对广地龙的DNA分子鉴定研究  

Identification of Pheretima aspergillum by COⅠ and 16S rRNA with DNA Molecular Marker Methods

作　　者：韦健红[1] 李薇[1] 吴文如[1] 喻良文[1] 

机构地区：[1]广州中医药大学中药学院,广州510006

出　　处：《中国药房》2012年第35期3274-3278,共5页China Pharmacy

基　　金：广东省科技计划项目(2009B060700006);广州市科技计划项目(2010Y1-C981)

摘　　要：目的:建立一种快速、准确和标准化的广地龙DNA分子标记鉴别方法。方法:测定了5个不同居群广地龙的线粒体细胞色素酶亚单位(CO)Ⅰ和16S rRNA基因序列,采用CodonCode Aligner进行序列拼接,通过下载GenBank地龙原动物的COⅠ与16S rRNA序列,采用MEGA4.1计算广地龙及其伪品地龙的种内、种间的K2P遗传距离,并基于K2P模型构建NJ和MP树。结果:COⅠ变异位点、信息位点均高于16SrRNA,COⅠ基因无插入和缺失,16S rRNA存在4个插入和缺失。COⅠ和16S rRNA序列种间遗传距离均明显大于种内,COⅠ和16S rRNA基因均能将广地龙从其他地龙或蚯蚓物种鉴别开来。结论:获得的广地龙COⅠ和16S rRNA序列可为动物性中药材地龙的分子水平鉴定提供参考,为动物性中药材DNA条形码数据库积累了相关信息数据。OBJECTIVE： To establish a rapid, accurate and standardized DNA molecular marker method for the identification of Pheretima aspergillum. METHODS： The CO Ⅰand 16S rRNA gene sequences of P. aspergillum from 5 different populations were determined using CodonCode Aligner sequence assembly. In addition, the CO Ⅰ and 16S rRNA gene sequences of P aspergil- lum were downloaded from GenBank, and intraspecific and interspecific K2P genetic distance between P aspergillum and counter- feit were calculated with MEGA 4.1 to construct NJ and MP phylogenetic tree. RESULTS： The variation sites and informa- tion sites of CO I were higher than those of 16S rRNA. There were no insertions and deletions of COⅠ , and there were 4 insertions and deletions of 16S rRNA. The interspecific genetic distances of CO Ⅰ and 16S rRNA sequences were significantly greater than in- traspecific ones, and COⅠ and 16S rRNA gene can be identified from the other earthworm species. CONCLUSION： The CO Ⅰand 16S rRNA gene can provide reference for the identification of the animal Chinese medicine at the molecular level, and accumulate information for DNA barcode of animal Chinese medicine.

关 键 词：地龙 线粒体细胞色素酶亚单位Ⅰ 16S RRNA 分子鉴定 

分 类 号：R93[医药卫生—生药学] R282.73[医药卫生—药学]