金钱鱼毒腺cDNA表达文库的构建及EST序列分析

以金钱鱼 (Scatophagusargus)毒腺为出发材料 ,构建以真核表达载体pcDNA3 0为基础的金钱鱼毒腺cDNA表达文库 .应用SMARTTM cDNALibraryConstruction技术和生物信息学分析等方法 ,通过对文库克隆的序列测定和初步生物信息学分析 ,获得了 2 0 1个金钱鱼新表达序列标签 (ESTs) ,其中已确定全长cDNA的克隆有 2 7个 ,包括多个核糖体大小亚基蛋白 (ribosomalprotein)、细胞凋亡相关蛋白 (programmedcelldeath 10 ) ,G蛋白信号调控子 (Gproteinsignalingregulator)、胸腺素(thymosinbeta 4 )、延伸因子 (translationelongationfactor 1 alpha)和泛素 (ubiquitin)等 .金钱鱼毒腺cDNA表达文库的成功构建 ,为研究金钱鱼毒腺的活性组分及其作用机制奠定了基础 ,也是分离新基因不可多得的重要资源

Construction of cDNA Expression Library from the Venom of Scatophagus argus and EST Analysis

A cDNA expression library from the venom of Scatophagus argus was constructed into pcDNA3.0 eukarytical expression plasmid. SMART TM cDNA Library Construction protocol was used for cDNA library construction and bioinformatics analysis was carried out. 201 novel ESTs (expressed sequence tags) were obtained from 250 clones in the library, of which there were 27 full length clones, including ribosomal protein gene, programmed cell death 10 gene, thymosin beta 4 gene, translation elongation factor 1 alpha gene, ubiquitin gene and so on. Most of these genes were reported for the first time in Scatophagus argus. Further studies on those genes and large scale sequencing of the library may be helpful to elucidate the components of Scatophagus argus venom and understand the function of those proteins at molecular level.