巨型艾美耳球虫孢子化卵囊cDNA文库的构建  

Construction of the cDNA Library of Sporulated Oocysts of E.maxima

作　　者：张建哲[1] 韩红玉[2] 姜连连[2] 董辉[2] 赵其平[2] 卞庆松[2] 闫晓菲[2] 黄兵[2] 

机构地区：[1]上海师范大学生命与环境科学学院,上海200234 [2]中国农业科学院上海兽医研究所/农业部动物寄生虫学重点开放实验室,上海200232

出　　处：《生物技术通报》2008年第2期195-198,共4页Biotechnology Bulletin

基　　金：国家科技基础条件平台建设项目(2005DKA21205-4)

摘　　要：为构建巨型艾美耳球虫(Eimeria maxima)孢子化卵囊cDNA表达文库,从E.maxima孢子化卵囊中提取总RNA,以总RNA为模板、λTriplex2TM为载体,利用SMARTTM cDNA文库构建试剂盒构建全长cDNA表达文库。经测定构建的E.maxima cDNA表达文库的原始文库容量为1×106 pfu/ml,扩增后的文库滴度为5×1010 pfu/ml,重组率为95%,插入片断主要集中在0.5～1kb之间。根据已知序列设计引物,能从文库中扩增出编码E.maxima免疫球蛋白重链结合蛋白的基因序列片段。结果表明所构建的E.maxima cDNA表达文库质量良好,为克隆、筛选E.maxima的功能性基因奠定了基础。To construct a full-length cDNA library of sporulated oocysts of Eimeria maxima,with the total RNA isolated from the sporulated oocysts of E. maxima as template and λTriplex2^TM as vector,a full-length cDNA library was constructed following the procedures provided by the SMARTTM cDNA library construction kit. The titer of unamplified library is 1 × 10^6 pfu/ml and the titer of. amplified library is 5 × 10^33 pfu/ml. The percentage of recombinant clones is 95%. E. maxima gene for immunoglobulin heaW chain binding protein can be amplified from the constructed library. This library has high quality and can be qualified for new gene selected.

关 键 词：球虫 巨型艾美耳球虫 CDNA文库 SMART技术 

分 类 号：S852.7[农业科学—基础兽医学] Q78[农业科学—兽医学]