欧猬迭宫绦虫全长cDNA文库构建及鉴定

Construction and Identification of a Full-Length cDNA Library from Spirometra erinaceieuropaei

应用SMART方法成功构建欧猬迭宫绦虫成虫全长cDNA文库,文库的重组率为95.5%,库容量为1.06×106;平均插入片段长度约为1.4kb。选取48个随机阳性克隆测序,去除<450bp序列后获得36个有效表达序列标签(EST),平均长度为674bp,其中单基因簇(unigene)比例为58.3%(21/36);26条具有同源性序列的EST中15条有或可能有5′末端,全长性比率为57.7%(15/26)。文库质量良好,可用于大规模EST测序。

The full-length pBluescript Ⅱ SK cDNA library of adult Spirometra erinaceieuropaei was constructed by using the SMART method. Data showed that 95.5% of the library was recombinant and the titer of the library was 1.06×106. The average insert size of the library was about 1.4 kb. Forty-eight randomly selected clones were sequenced. A set of 36 effective expressed sequence tags （ESTs） with the average size of 674 bp was obtained after excluding clones shorter than 450 bp. The unigenes occupied 58.3% of the 36 ESTs. The rate of full-length cDNAs were 57.7% （15/26）. The high-quality of full-length cDNA library could be used for large scale EST sequencing.