作者：叶庆亮,江东,彭爱红
作者单位：中国农业科学院柑桔研究所
摘要：以‘岩溪晚芦’椪柑(Citrus reticulata Blanco)成熟果实果皮和果肉为材料,采用抑制性差减杂交技术成功构建了果皮(tester)与果肉(driver)的差减cDNA文库。结果显示:cDNA克隆外源插入片段大小介于100～500bp之间。成功对411个克隆进行了测序,测序结果经与NCBI基因库中序列进行比较,377个EST找到了同源序列,它们分属于126个基因,涉及到色素合成、能量代谢、初级代谢、次生代谢、抗逆防御、蛋白代谢、信号转导、香味形成、果皮软化(衰老)、精油合成、抗氧化等代谢途径和生理生化过程。还有一些基因涉及到三价铁螯合物、反转录转座子gag蛋白、查尔酮合成、硼离子转运、儿茶酚氧位甲基转移酶等功能。功能已知且没有重复的EST共有63个,占总数的49.2%。有5个基因共62个克隆虽在Blast搜索时找到了较高的同源性序列,但功能目前尚未明确。另有34个基因未搜索到同源序列,可能为新发现的基因。
关键词：柑橘,抑制性差减杂交(SSH),cDNA文库,表达序列标签,克隆,差异表达基因
Title: Construction of a Subtractive cDNA Library Between Fruit Epicarp and Pulp of Ponkan Mandarin and Primary Analyses of the Sequences
Authors: YE Qing-liang,JIANG Dong,PENG Ai-hong
Abstract: A cDNA library was successfully constructed via suppression subtractive hybridization between fruit epicarp(experimental group,tester)and pulp(control group,the driver)of Ponkan(Citrus reticulata Blanco).Results showed that most of the clones had inserting fragments which length ranged from 100 bp to 500 bp.Four hundred and eleven ESTs were successfully sequenced and searched against NCBI GenBank,the results indicated that 377 ESTs had significant similarities with known genes.All the ESTs were divided into 126 genes according to the gene annotation.These ESTs were mainly involved in pigment synthesis,energy metabolism,primary metabolism,secondary metabolism,defense against stresses,signal transduction,transcription,flavor formation,oil synthesis,peel softening,antioxidant metabolism etc.There were 63 uni-ESTs which accounted for 49.2% of the total ESTs and another 34 ESTs had no matching homologous sequences found.
Key words: citrus； suppression subtractive hybridization(SSH)； cDNA library； expressed sequence tag； clone； di
文献注录：叶庆亮,江东,彭爱红. ‘岩溪晚芦’椪柑果皮与果肉差减cDNA文库的构建及初步分析 [J]. 园艺学报. 2009, 07: 967-974.
基金: 国家科技支撑计划项目(2006BAD13B06-1-4);科技部‘973’重大基础研究前期研究专项(2005CCA02900;CSTC,2007BB1343);市自然科学基金项目(SCTC2005BA1010)
报/刊名：《园艺学报》，发表于2009 / 第 07 期。
文献类型： [J] （文献级别：核心刊物）
页码：第 967-974 页 / 共8 页