芝麻全长cDNA文库的构建及序列分析  

Construction and Sequence Analysis of a Full-length cDNA Library for Sesame

作　　者：魏利斌[1] 苗红梅[1] 张体德[1] 李春[1] 张海洋[1] 

机构地区：[1]河南省农业科学院芝麻研究中心,郑州450002

出　　处：《分子植物育种》2013年第5期617-623,共7页Molecular Plant Breeding

基　　金：国家重点基础研究发展计划973项目(2011CB109304);现代农业产业技术体系建设专项(CARS-15)共同资助

摘　　要：本研究采用Gateway建库技术构建了中国芝麻主栽品种豫芝11号的植株生长发育过程中不同组织的全长cDNA文库（SiFcDNA）。该全长cDNA文库容为5．6×10^6，文库滴度为1．12×10^6cfu／mL。部分克隆检测显示，文库cDNA片段长度集中于1．0～3．0kb，重组率为96．88％，全长cDNA比例为88％。随机挑取1152个单克隆进行5′端EST测序，共获高质量有效EST序列1088条，拼接得到单一基因序列867条，其中46条单一基因在NCBI数据库中没有查到相应的同源序列，可能为芝麻生长发育过程特有的新基因；所获单一基因序列被划分为26个GO功能亚类。此外，从获得的Unigene序列中挖掘出了173个SSR，分布频率为4．78kb／SSR；AG／CT类型SSR出现次数最多，占总SSR的19．08％。高质量全长SiFcDNA文库的构建为深入开展芝麻功能基因组学研究奠定基础。A sesame full-length cDNA library （SiFcDNA） was constructed based on plant growth and development with different tissues of Yuzhi No. 11 for a major cultivated cultivar in China using the gateway cloning technology. The cDNA library capacity reached a total of 5.6×10^6 clones and had the high titer of 1.12×10^6 cfu/mL. Analysis results of selected clones indicated that main cDNA fragments ranged from 1.0-3.0 kb with a recombination of 96.88%, and the ratio of full-length cDNAs was 88%. 1 152 clones were selected randomly from the SiFcDNA library and sequenced using 5＇ end sequencing method. A total of 1 088 ESTs were obtained and 867 unigenes were assembled. Bioinformatics analysis showed that 46 unigenes among the above sequences had no homolog matches in NCBI database and probably belonged to the new specific genes for sesame growth and development. GO functional analysis reflected that 867 unigenes with the functional annotations were involved in 26 various functions. A total of 173 SSRs were detected from these unigenes with the distribution frequency of 4.78 kb per one EST-SSR. Among the SSRs, the di-nucleotide type of AG/TC was the most abundant with the high frequency of 19.08%. Construction of the SiFcDNA library with high quality provided the material basis for further functional genomics research in sesame.

关 键 词：芝麻 全长CDNA文库 生物信息学 EST-SSR 

分 类 号：S565.3[农业科学—作物学]