千里光全长cDNA文库的构建及分析  

Construction of a full-length cDNA library for Senecio scandens

作　　者：平军娇[1] 张珍[1] 蔡振锋[1] 汤贤春[1] 钱刚[1] 

机构地区：[1]遵义医学院细胞生物学与遗传学教研室,贵州遵义563099

出　　处：《中草药》2012年第3期557-561,共5页Chinese Traditional and Herbal Drugs

基　　金：贵州省优秀科技教育人才省长基金项目(黔省专合字2008-61号)

摘　　要：目的构建千里光全长cDNA文库,以期研究千里光的功能基因组学信息,为克隆药理学性状相关的功能基因提供数据资源。方法 Trizol法提取千里光叶片总RNA,通过SMART(switching mechanism at 5’end of RNA transcript)构建全长cDNA文库,随机挑取600个单克隆测序分析文库滴度、全长率及冗余率,得到的EST序列进行Blast分析(NR、NT、Swiss-Prot、KEGG)及COG功能分类。结果文库的库容为4.3×106cfu/mL,插入片段大小平均1.7 kb,文库重组率96.35%,全长率58.24%,冗余率10.88%;获得524条全长EST序列,含有467条独立基因(unigenes),其中5条序列与千里光次生代谢产物的合成、运输与代谢有关。结论经检测,SMART技术成功构建了千里光全长cDNA文库,该文库可用于千里光功能基因组鉴定、新基因筛选及次生代谢产物生物合成的表达调控研究。Objective To establish the phylogenic map of plants in Epimedium L.based on PCR-RFLP data.Methods Seventeen species from Epimedium L.native to Sichuan and Guizhou provinces etc.were studied by means of PCR-RFLP Marker.Results Among the eight PCR-RFLP Markers,seven（87.5%） could produce one distinct band with no polymorphism detected by direct electrophoresis in 2% agrose gels.After the amplified products were digested by 12 restriction enzymes,a total of 129 bands were detected in 84 PCR-RFLP primer/enzyme combinations.And 44 polymorphic bands accounting for 34.1% have polymorphism.The genetic similarity coefficient（Gs） among 17 species from Epimedium L.showed the range from 0.550 to 0.988,with the mean of 0.821.Conclusion It is clear that there are some relationships between the genetic property and its geographic distribution.

关 键 词：千里光 全长CDNA文库 SMART EST 基因组学 

分 类 号：R282.12[医药卫生—中药学]