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.pdfVIP

中国农业科学 2007,40(7):1331-1336 Scientia Agricultura Sinica 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 赵光耀，孔秀英，贾继增 （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） 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6 973条和叶EST 6 616 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。 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 Full-length cDNA Libraries Construction from Ae. tauschii Root and Shoot and EST Annotation and Comparative Analysis ZHAO Guang-yao, KONG Xiu-ying, JIA Ji-zeng (Key Laboratory of Crop Germplasm Biotechnology, Ministry of Agriculture, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences ; The National Key Facility for Crop Gene Resources and Genetic Improvement, Beijing 100081) Abstract: 【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,