欧李叶片全长cDNA文库的构建和部分克隆的序列分析  

Construction and Sequence Analysis of Partial Clones of a Full-Length cDNA LibraryofChinese Dwarf CherryLeaves

作　　者：高小丽[1] 李芳[1] 岳鹏[1] 李天红[1] 

机构地区：[1]中国农业大学农学与生物技术学院果树系,北京市果树逆境生理与分子生物学重点开放实验室,北京100193

出　　处：《农业生物技术学报》2010年第1期156-162,共7页Journal of Agricultural Biotechnology

基　　金：新世纪人才支持计划(No.NCET-06-0108);国家"十一五"科技支撑计划(No.2007BAD36B02);国家自然科学基金(30871696)共同资助

摘　　要：采用SMART技术,构建了欧李(Prunus humilis Bunge)品系T8-1叶片全长cDNA文库,原始文库滴度达6.17×107pfu/mL,库容达3.7×107,重组率达95%,平均插入片段大小约为1.2kb。随机挑取400个单克隆进行5'端测序,共获得364个欧李ESTs。绝大部分ESTs的长度在500～1300bp之间,平均长度为877bp,其中具有完整ORF结构的序列有221条,占60.71%。通过与NCBI等非冗余核酸数据库和蛋白质数据库进行比对、查询和注释,获得已知功能基因或具推测功能的基因181个(339个ESTs),相似性较低的未鉴定基因5个(9个ESTs),新基因14个(16个ESTs)。A full-length cDNA library of chinese dwarf cherry（Prunus humilis Bunge） leaves was constructed by using λTriplEx2 vector according to the SMART cDNA library kit protocol.The titer of primary cDNA library was 6.17×107 pfu/mL containing 3.7×107 clones and the rate of recombinant was 95%.The average size of insert fragments was about 1.2 kb.400 clones from the cDNA library were sequenced and the first 364 valid chinese dwarf cherry ESTs were generated.Most of the ESTs were between 500～1 300 bp with an average of 877 bp and the rate of sequences containing whole open reading frame was 60.71%.181 known or hypothetical functional genes 5 unidentified genes and 14 novel genes were annotated by Blastx and Blastn searches against the NCBI non-redundant protein and nucleotide databases.

关 键 词：欧李 全长CDNA文库 ESTS 

分 类 号：S435.621.2[农业科学—农业昆虫与害虫防治]