毒害艾美耳球虫卵囊壁蛋白的SDS-PAGE及Western blot分析  

Analysis of Western blot and SDS-PAGE for oocyst wall proteins of Eimeria necatrix

作　　者：杜彩娟 查夕馨 黄晓星[1] 刘丹丹[1] 许金俊[1] 陶建平[1] 

机构地区：[1]扬州大学兽医学院江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《中国兽医学报》2017年第5期854-858,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31472181);高等学校博士学科点专项科研基金资助项目(20123250110002);大学生实践创新训练资助项目(201411117032Z);江苏省高校优势学科建设工程资助项目

摘　　要：利用聚丙烯酰胺凝胶(SDS-PAGE)电泳技术,分析了毒害艾美耳球虫未孢子化卵囊壁可溶性蛋白。用鼠抗毒害艾美耳球虫重组配子体蛋白rEnGAM56和rEnGAM59多克隆抗体,对卵囊壁可溶性蛋白进行了Western blot分析。结果显示,至少有24条较清晰的电泳条带,其中6条为相对明显的主带,其相对分子质量分别为37 000,35 000,34 000,20 000,14 000和12 000。Western blot分析显示,鼠抗rEnGAM59和rEnGAM56多抗均能识别1条条带,前者的相对分子质量约14 000,后者约30 000。推测30 000和14 000卵囊壁蛋白分别来自毒害艾美耳球虫配子体蛋白EnGAM56和EnGAM59。The soluble protein profiles of unsporulated oocyst wall proteins of Eimeria necatrix were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis technique （SDS- PAGE）. The antigens of the proteins were respectively detected by the anti-rEnGAM59 and anti- rEnGAM56 polyelonal antibodies using western blot technique. The results showed that there were at least twenty-four protein bands,including six predominant proteins with 37 000,35 000,34 000, 20 000,14 000 and 12 000,respectively. Only one protein band was recognized by the two antibod- ies,respectively. The molecular weight was 14 000 by the anti-rEnGAM59 antibody,and 30 000 by the anti-rEnGAM59 antibody. It would suggest that the 30 000 proteins were derived from GAM56 and the 14 000 protein was derived from GAM59.

关 键 词：毒害艾美耳球虫 卵囊壁蛋白 SDS-PAGE WESTERN BLOT 

分 类 号：S852.7[农业科学—基础兽医学]