鳜肌肉组织cDNA文库构建及其ESTs分析

以鳜(Siniperca chutasi)肌肉为实验材料,采用非均一化引物定向克隆技术构建了鳜肌肉组织cDNA文库,并进行大规模EST测序和序列分析。结果显示,cDNA文库的库容量为2.3×105,重组率达96.5%,平均插入片段长度为1.2 kb。随机挑选5456个克隆,成功测序5191个,其中高质量序列5063个,达97.5%。利用PHRAP程序聚类拼接后,得到1625条非冗余序列,包括443个重叠群(Contigs),1182个单拷贝(Singlets)。使用BLAST软件将这些序列同GenBank等数据库进行比对、注释,发现1625个非冗余序列与鳜肌肉结构和发生相关,而其中991条序列与NCBI数据库中的其它物种已知序列存在显著的相似性,剩余的634条非冗余序列(占所有非冗余序列的39%)为在鳜中新发现的未找到同源序列的序列。740个基因为能够注释到Gene ontology(Go)中的序列数目。

Construction of muscle cDNA library and analysis of some expressed sequence tag sequences of Siniperca chutasi

The cDNA library of Siniperca chutasi′s muscle was constructed by non-normalizad cDNA library construction techniques.The library capacity was 2.3×105,the recombination frequency was 96.5%,and the average size of inserts was 1.2 kb.Of total 5456 sequencing reactions,5191 were successfully sequenced,in which 5063 were in high-quality,which was 97.5%.1625 non-repetitive sequences including 443 contigs and 1182 singlets were analyzed with PHRAP software.The 1625 unique sequences were functionally related to muscle structure and myogenesis as searched from NCBI databases.991 of 1625 unique sequences were found to have high similarity to other fish species and the rest of 634 sequences were unique in S.chutasi accounting for 39 percent of the whole.Further,740 functional known genes were categorized by Gene ontology(GO).