鸡堆型艾美耳球虫孢子化卵囊cDNA文库的免疫学筛选  

Immunological Screening of cDNA Expression Libraries in Sporulated Oocysts of Eimeria acervulina

作　　者：阎晓菲[1] 韩红玉[2] 黄兵[2] 岳城[3] 赵其平[2] 姜连连[2] 董辉[2] 

机构地区：[1]新疆农业大学科学技术学院,新疆乌鲁木齐830000 [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实验室,上海200241 [3]新疆农业大学动物医学学院,新疆乌鲁木齐830052

出　　处：《安徽农业科学》2014年第14期4363-4364,4454,共3页Journal of Anhui Agricultural Sciences

基　　金：国家科技基础条件平台建设项目(2005DKA21205-4)

摘　　要：[目的]寻找鸡球虫保护性抗原基因,为研究基因功能和研制抗鸡球虫疫苗奠定基础.[方法]将纯化的堆型艾美耳球虫孢子化卵囊于兔皮内多点注射,制备堆型艾美耳球虫抗血清,用间接酶联免疫吸附试验（ELISA）检测其效价,进行免疫学筛选.利用PCR对获得的阳性克隆进行初步鉴定.[结果]间接酶联免疫吸附试验结果表明堆型艾美耳球虫抗血清检测结果呈阳性,初步筛选到的疑似阳性斑经3次复筛后获得6个阳性克隆,1～3号克隆约为1 000bp,4～6号克隆约为1 500 bp.[结论]对堆型艾美耳球虫孢子化卵囊cD-NA文库获得的阳性克隆成功进行了免疫学筛选.[Objective] The research aimed to find out the protective antigen gene of chicken coccidian and lay the foundation for studying the gene function and producing vaccine against chicken coccidiosis.[Method] The purified sporulated oocysts of Eimeria acervulina was injected in many points of rabbit skins to prepare the antiserum of E.acervulina.And its titer was detected by ELISA to make immunological screening.And the obtained positive clone was identified by PCR.[Result] The results of ELISA showed that the antiserum detection results of E.acervulina was positive.After preliminary screening,suspected positive plaques were obtained,then 6 positive clones were obtained after three times of second screening.The size of 1-3 clone were about 1 000 bp and that of 4-6 clone were about 1 500 bp.[Conclusion] The immunological screening of the positive clones obtained from cDNA library of sporulated oocysts of E.acervulina was made.

关 键 词：鸡 球虫 堆型艾美耳球虫 CDNA文库 免疫学筛选 

分 类 号：S821[农业科学—畜牧学]