水滴伪康纤虫全长cDNA文库的构建及EST分析  

Full-length cDNA Library and EST Analysis on Pesudocohnilembus persalinus

作　　者：林能锋[1] 曾红[2] 

机构地区：[1]福建省农业科学院生物技术研究所,福建福州350003 [2]福建师范大学生命科学学院,福建福州350117

出　　处：《福建农业学报》2017年第9期921-925,共5页Fujian Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31201698);福建省自然科学基金项目(2013J01120);福建省科技计划项目--省属公益类科研院所基本科研专项(2015R1019-11)

摘　　要：水滴伪康纤虫为海水养殖鱼类的重要危害性寄生虫。采用SMART技术构建营养期水滴伪康纤虫全长cDNA文库,初始文库滴度3.3×106pfu·mL^(-1),文库容量约为1.5×107 pfu,重组率97%。随机挑取24个克隆,经PCR检测插入的片段长度在0.3~2.0kb,平均长度>500bp。挑选文库中的1 200个克隆进行表达序列标签测序,得到1 032条高质量的EST,与NCBI数据库进行比对后,获得292个同源基因,其中包含190条单拷贝EST和102个重叠群,冗余度为71.7%。有215条EST在Tetrahymena thermophila,Paramecium tetraurelia,Ichthyophthirius multifiliis,Cryptocaryon irritans,Oxytricha trifallax等自由生活及寄生种类的纤毛虫的基因组中比对到同源基因,其中有134条EST具有功能注释,其余为未知功能的假定蛋白基因。水滴伪康纤虫cDNA文库的构建及EST测序的完成,为进一步发现和研究水滴伪康纤虫的功能基因奠定了基础。Pesudocohnilembus persalinus is a harmful parasite found in marine fish.Using SMART technology,a fulllength cDNA library of P.persalinus was constructed.Titer of the primary library was 3.3×106 cfu·mL^（-1）,and the library capacity was 1.5×107 cfu with a recombination rate of 97%.A PCR amplification on 24 random clones revealed that the inserted cDNA fragments ranged from 300-2 000 bp,with an average length over 500 bp.Subsequently,1 200 clones were selected from the library for EST sequencing.As a result,1 032 high-quality ESTs were obtained and assembled into 292 unigenes including 102 contigs and 190 unique ESTs with a redundancy rate of 71.7%.In a blast analysis against NCBI database,215 of the unigenes were homologous to the corresponding genes of ciliates with free-living and parasitic lifestyle,such as Tetrahymena thermophila,Paramecium tetraurelia,Ichthyophthirius multifiliis,Cryptocaryon irritans,Oxytricha trifallax,etc.But,only 134 ESTs had been annotated.The remainders were hypothetical protein genes.The results obtained from this study would substantially facilitate the molecular cloning and characterization of the functional genes in P.persalinus.

关 键 词：水滴伪康纤虫 CDNA文库 表达序列标签 

分 类 号：Q959[生物学—动物学]