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双价工程菌K99和F41菌毛抗原的提纯、特征分析及免疫效果观察

Purification and Properties of Recombinant K99 and F41 Fimbriae Antigens From Engineered Strain

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【作者】 钟声李丰生芮贤良吕学洗王庆元黄培堂

【Author】 Zhong,Sheng Li,Feng-sheng Rui,Xian-liang Lu,Xue-xian Wang,Qing-ynan Huang,Pei-tang(Molecular Genetics Center,Institute of Biotechnology,Beijing 100850)

【机构】 佳木斯医学院生物教研室军事医学科学院生物工程研究所军事医学科学院生物工程研究所 佳木斯 154002北京 100850佳木斯 154002北京 100850

【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。

【Abstract】 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】 K99菌毛F41菌毛免疫原性
【Key words】 K99 fimbriaeF41 fimbriaePurification
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