粗山羊草(Ae.tauschii)幼苗和根全长cDNA文库构建及其EST注释与比较分析  

Full-length cDNA Libraries Construction from Ae.tauschii Root and Shoot and EST Annotation and Comparative Analysis

作　　者：赵光耀[1] 孔秀英[1] 贾继增[1] 

机构地区：[1]中国农业科学院作物科学研究所/农业部作物种质资源与生物技术重点开放实验室国家农作物基因资源与基因改良重大科学工程,北京100081

出　　处：《中国农业科学》2007年第7期1331-1336,共6页Scientia Agricultura Sinica

基　　金：农业部"948"项目(2005-Z47);"973"国家重大基础研究发展规划(2004CB117200)

摘　　要：【目的】构建小麦D基因组供体粗山羊草根和叶的全长cDNA文库,探讨利用生物信息学方法分析发掘组织特异性表达基因的可行性。【方法】本研究利用Cap-trapper方法构建全长cDNA文库,利用高通量测序技术获得大批高质量EST序列,在基于Linux系统的本地化生物信息学分析平台上对上述EST进行了电子注释分析。【结果】成功构建了粗山羊草根和叶的全长cDNA文库,测序获得根EST序列6973条和叶EST6616条。利用本地化数据分析平台对其进行功能注释。利用GO-Diff软件,发现了两个文库间表达基因的功能差异,找到组织间表达差异显著的基因和组织内特异性表达的基因。【结论】利用构建的全长cDNA测序获得大量EST序列,通过生物信息学方法挖掘差异表达或特异性表达基因的方法是可行的。[Objective] Two full-length cDNA libraries from Ae. tauschii shoot and root mRNA were constructed applying Cap-trapper method. Two sets of EST data were generated and differentially expressed genes were detected and ananlyzed via a bioinformatics program. [Method] The Cap-trapper method was optimized by reporter＇s lab and a localized bioinformatics analysis tool based on a Linux operation system was tested and applied. [Result] Two high quality full-length cDNA libraries were constructed successfully from Ae. tauschii cv. Y2282. 6 616 ESTs and 6 973 ESTs were obtained from the two libraries respectively. EST function was annotated against public databases such as protein, nucleotide and Triticum and Aegilops EST databases from GenBank etc. By using results from Gene Ontology catalogue obtained by software GoPipe2 and expression profile from Phrap assembling, some differentially or specifically expressed ESTs were identified in the two libraries. The results give suggestion in molecular basis of different plant organ functions performed. [Conclusion] The Cap-trapper method proved to be excellent in full-length cDNA library construction from Ae. tauschii. EST analysis was also competent in searching for differentialy or specifically expressed genes in various tissues or physiological states with further bioinformatics help.

关 键 词：粗山羊草 表达序列标签 全长CDNA文库 功能注释 生物信息学 

分 类 号：S512.1[农业科学—作物学]