观音竹盐胁迫均一化全长cDNA文库的构建与EST测序分析

摘要

将DSN(duplex-specific nuclease)均一化技术与SMART(switching mechanism at 5'end of RNA transcript)建库技术相结合,构建观音竹(Bambusa multiplex var.riviereorum)盐胁迫幼叶的均一化cDNA文库.经检测该原始文库滴度为1.7×106cfu·mL-1,文库重组率达97％,插入片段的平均长度为1.5 kb.同时对文库中随机的600个单克隆进行测序,获得543个非重复序列,包括30个片段重叠群和513个单基因;文库冗余率为5.4％.表明观音竹盐胁迫幼叶均一化全长cDNA文库构建成功.%A normalized full-length cDNA library of the young leaf of Bambusa multiplex var. Riviereorum was established by themethod of DSN (duplex specific nuclease) normalization and SMART (switching mechanism at 5'end of RNA transcript). The titerof unamplified cDNA library was about 1.7 x 106 cfu · mL-1. The average size of cDNA inserts was 1500 bp with the recombinationrate of 97%. Meanwhile a total of 543 unigenes were attained, including 30 contigs and 513 singltons by sequencing and analyzing600 cDNA clones of normalized cDNA library. These results indicated that the normalized full-length cDNA library was establishedsuccessfully.