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

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.