巨型艾美球虫株间差异表达基因消减文库的构建和分析  

Construction and analysis of subtractive cDNA libraries of differential immunogenicity strains of Eimeria maxima by suppression subtractive hybridization

作　　者：孙文瑶[1] 许金俊[1] 陶建平[1] 吴海清[1] 李金贵[1] 彭昊[1] 张艳[1] 姜慧婕[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2009年第1期13-17,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：江苏省自然科学基金资助项目(B2007079);江苏省高校自然科学基金资助项目(VK051095)

摘　　要：为分离鉴定巨型艾美球虫上海株与南通株间免疫原性的差异表达基因,分别以巨型艾美球虫上海株孢子囊cDNA为驱动组,以南通株孢子囊cDNA为试验组,或相反,利用抑制消减杂交(SSH)方法,通过2轮杂交和2次PCR分别构建了2个抑制性消减文库。随机从2个消减文库中分别挑取96个克隆产物,经PCR鉴定上海株和南通株孢子囊消减文库的重组率分别为91%和94%,差异基因片段大小为250～1 000 bp。从每个文库中随机挑取30个阳性克隆测序,并进行同源性比较分析,结果表明:从上海株cDNA消减文库中获得了19个单一有效序列,其中15个为未知序列;从南通株cDNA消减文库中获得了16个单一有效序列,其中13个为未知序列。这些结果将为分离巨型艾美球虫新功能基因和进一步探索球虫抗原变异相关的分子机理奠定基础。In order to clone and identify differentially expressed genes in two immunologically distinct strains of Eimeria maxima,the two subtractive cDNA libraries of sporcysts of Nantong strain(NT) and Shanghai strain(SH) were constructed using the technique of suppression subtractive hybridization(SSH),respectively.PCR amplification revealed that the SH strain and the NT strain subtractive cDNA libraries contained approximated 91%and 94%recombinant clones,respectively,and the lengths were 250—1000 bp.Ninety-six po...

关 键 词：巨型艾美球虫 免疫原性相异株 消减文库 构建 

分 类 号：S852.7[农业科学—基础兽医学]