El-Ashram Saeed, Yin Qing, Liu Hongbin, Al Nasr Ibrahim, Liu Xianyong, Suo Xun, Barta John
State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing, 100193, China.
National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China.
PLoS One. 2015 Dec 7;10(12):e0143232. doi: 10.1371/journal.pone.0143232. eCollection 2015.
Two immunologically distinct strains of E. maxima were examined in this study: the M6 strain and the Guelph strain. The differential expression between the sporozoites of the two strains of E. maxima was determined by image analysis of 100 μg of protein from each strain separated by standard one- and conventional two-dimensional polyacrylamide gel electrophoresis. In addition to differences in both molecular weight and the electrophoretic mobility, differences in the intensity of polypeptide bands for example, GS 136.4 and M6 169 were explored. Pooled gels were prepared from each strain. A representative 2D-PAGE gel spanning a non-linear pH range of 3-10 of E. maxima strain M6 consisted of approximately 694 polypeptide spots with about 67 (9.6%) of the polypeptide spots being unique relative to the other strain. E. maxima strain GS had about 696 discernable polypeptide spots with 69 spots (9.9%) that differed from those of the M6 strain. In-depth characterization of the variable polypeptide spots; unique polypeptide spots (absence or presence) and shared polypeptide spots with modifications may lead to novel vaccine target in the form of multi-component, multi-stage, multi-immunovariant strains, multi-species subunit vaccine, and diagnostic probe for E. maxima.
M6菌株和圭尔夫菌株。通过对每种菌株100μg蛋白质进行标准一维和传统二维聚丙烯酰胺凝胶电泳分离后的图像分析,确定了这两种巨型艾美耳球虫菌株子孢子之间的差异表达。除了分子量和电泳迁移率的差异外,还探讨了多肽条带强度的差异,例如GS 136.4和M6 169。从每种菌株制备合并凝胶。一张代表性的巨型艾美耳球虫M6菌株的二维聚丙烯酰胺凝胶电泳图谱,其非线性pH范围为3至10,大约由694个多肽斑点组成,其中约67个(9.6%)多肽斑点相对于另一种菌株是独特的。巨型艾美耳球虫GS菌株约有696个可分辨的多肽斑点,其中69个(9.9%)与M6菌株的不同。对可变多肽斑点进行深入表征;独特的多肽斑点(存在或不存在)以及有修饰的共享多肽斑点,可能会产生以多组分、多阶段、多免疫变体菌株、多物种亚单位疫苗形式的新型疫苗靶点,以及用于巨型艾美耳球虫的诊断探针。