亚热带植物科学Issue(2)：93-97,5.DOI:10.3969/j.issn.1009-7791.2014.02.001
德保苏铁珊瑚根均一化全长cDNA文库的构建和EST分析
Construction of Normalized Full-length cDNA Library for the Coralloid Roots of Cycas debaoensis and Analysis of its Expressed Sequence Tags
摘要
Abstract
A normalized full-length cDNA library was constructed with the coralloid roots of Cycas debaoensis by DSN (duplex-specific nuclease) normalization method combined with SMART (Switching Mechanism At 5′ end of the RNA Transcript) technique. The titer of original cDNA library was about 1.5×106 cfu·mL-1 and the average insertion size was about 1 kb with high recombination rate (97%). The 175 high quality expressed sequence tags (ESTs) were obtained from 192 randomly picked cDNA clones. Clusting and assembly of ESTs resulted in 165 unique sequences, consisting of 7 contigs and 158 singlets. Using a generic similarity threshold (score≥200, identity≥60), approximately 115 (66.1%) of all 174 clean ESTs matched corresponding homologous sequences according to BLAST analysis of the nr-nt (non-redundant protein and non-redundant nucleotide sequence) database in GenBank. The 33.9% (59) ESTs have no significant match. These results showed preliminarily that we successfully constructed a normalized full-length cDNA library of coralloid roots in C. debaoensis, which could serve as an abundant information platform for functional marker development and functional gene study.关键词
德保苏铁/珊瑚根/均一化/cDNA文库Key words
Cycas debaoensis/coralloid root/normalized/cDNA library分类
农业科技引用本文复制引用
王运华,陈庭,李楠..德保苏铁珊瑚根均一化全长cDNA文库的构建和EST分析[J].亚热带植物科学,2014,(2):93-97,5.基金项目
国家林业局“珍稀濒危物种野外救护与繁育”财政专项(2010)、深圳市仙湖植物园基金项目(2012) （2010）
