摘要
本文利用加热搅拌及Sephadex G-100凝胶过滤方法,从双价重组工程菌RRI(pMG611)中分离了重组K99和F41菌毛抗原。SDS-PAGE测定其分子量,重组K99和F41抗原亚单位分子量分别是17200和29800,与各自野生菌毛亚单位分子量相同。甘露糖抗性血凝试验(MRHA)性质与野生K99和F41抗原相似。双向扩散试验和Western blot分析证实其免疫学性质亦与野生菌毛抗原相似。重组K99和F41抗原的免疫原性较强,能够刺激家兔产生高效价的抗体出现。
The recombinant K99 and F41 fimbriae antigens expressed by E.coli RRI (pMG611) strain had been purified by heat treatment and then by Sephadex G-100 filtratiion.The molecular weight of their subunit were determined by SDS-PAGE.They are the same as those of the corresponding fimbriae antigens from wild strains.The MRHA properties of recombinant antigens are similar to those of wild strains.Immunological characteristics of the recombinant fimbriae were demonstrated by double immunodiffusion test and Western blot assay.They are also similar to those of wild fimbriae.The high titer of antibody of both K99 and F41 reached in rabbit showed that they had strong immunogenicity.
关键词
双价工程菌
抗原
免疫原性
K99 fimbriae
F41 fimbriae
Purification
