猪带绦虫六钩蚴cDNA文库的构建及免疫原基因的筛选与克隆  

The Construction and Screening of cDNA Expression Library of Oncosphere of Taenia solium and cloning of Immunogen Gene

作　　者：景志忠[1] 郭爱疆[1] 海岗[1] 宗瑞谦[1] 才学鹏[1] 

机构地区：[1]中国农业科学院兰州兽医研究所农业部畜禽病毒病重点开放实验室,兰州730046

出　　处：《畜牧兽医学报》2005年第4期391-396,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重大基础研究"973"项目(G1999011906)

摘　　要：研究避开庞大的猪带绦虫基因组DNA,以构建六钩蚴发育阶段的cDNA表达文库为策略,利用pSPORT1质粒表达载体首次成功构建了表达文库。经库容量、重组率和代表性测定,库容量达5.0×106,重组率100%,用特异性引物能扩增出六钩蚴发育阶段10ku基因、18ku基因和45W基因家族的A型、B型、C型转录本,说明文库质量高,代表性强。应用快速筛选质粒表达文库和克隆免疫原基因的技术,成功地筛选和克隆到TsO1基因,其完整阅读框架(ORF)为393bp。将其登录到GenBank(AY327451),经BLAST分析,证实是猪带绦虫六钩蚴发育阶段的1个新免疫原基因。The cDNA expression library of the Oncosphere development stage was constructed in order to avoid involved in large genome of Teania solium. The results showed that the transformation efficacy and recombinant rate of library were 5.0×10~6 and 100%, respectively. The genes of 10 ku, 18 ku and A, B, C types of 45W were cloned from the cDNA library, which approved the library possessed a good quality and high representation. The library was screened with antiserum, and gene of TsO1 was cloned and sequenced. ORF size of TsO1 was 393 bp. The sequance of TsO1 was submited to GenBank, and the number AY327451 was given. It has been proved the TsO1 is a novel gene of the Oncosphere development stage by analysis of BLAST.

关 键 词：猪带绦虫 六钩蚴 免疫原 cDNA文库 克隆 构建 CDNA表达文库 基因组DNA 发育阶段 质粒表达载体 特异性引物 BLAST 基因家族 快速筛选 库容量 代表性 重组率 转录本 

分 类 号：S858.28[农业科学—临床兽医学] S852.734[农业科学—兽医学]