东亚三角涡虫cDNA文库的构建及EST初步分析

Construction of cDNA Library and Trial EST Analysis from Planaria(Dujesia japonica)

为了快速高效地分离鉴定东亚三角涡虫的发育和再生相关基因,以总RNA为模板,借助CreatorTM SMAR-TTM cDNA Library Construction Kit和Advantage 2 PCR Kit成功构建了东亚三角涡虫的cDNA文库。经测定,cDNA文库原始库容为1.22×105个独立克隆,重组率超过98%,插入片段平均大小为900bp。从文库中随机挑选重组克隆测序,并在NCBI数据库中比对,结果获得208个rRNA基因,148个编码蛋白基因,78个染色体片段基因。该研究为我国涡虫发育和再生的深入研究奠定了坚实的分子基础。

In order to speedily and effectively separate and identify the developmental and regenerative gene from Dujesia japonica, the total RNA as template were constructed the cDNA library, according to SMARTTM cDNA Library Construction Kit and Advantage 2 PCR Kit. The unamplified cDNA library contained 1.22 × 10^5 independent clones in capability. The recombination rate was more than 98%, The average inserted cDNA fragments were about 900 bp, with the majority ranging from 500 to 1500 bp, From the library, some DNA fragments was randomly selected to sequence and blast in NCBI database on line. The results showed that it contained 208 rRNA genes, 148 encoding genes and 78 genes in the chromosome. The results have established a solid molecular basis for the deep study of development and regeneration in planarian.