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来自奥斯特他线虫成虫的一种类珠蛋白抗原的分离、特性鉴定及免疫定位

Isolation, characterization and immunolocalization of a globin-like antigen from Ostertagia ostertagi adults.

作者信息

de Graaf D C, Berghen P, Moens L, de Marez T M, Raes S, Blaxter M L, Vercruysse J

机构信息

Department of Parasitology, University of Gent, Merelbeke, Belgium.

出版信息

Parasitology. 1996 Jul;113 ( Pt 1):63-9. doi: 10.1017/s0031182000066282.

Abstract

Western blot analysis using an anti-globin rabbit serum Rb94 revealed a major band of 17 kDa in extracts of Ostertagia ostertagi adults and 4th-stage larvae. The adult stage globin-like antigen (OoAdGlb) was purified from total worm extracts by liquid chromatography. The protein has an estimated molecular mass of 36 kDa under non-reducing conditions, suggesting a dimeric structure containing 2 non-covalently linked 17 kDa monomers. Tryptic peptides were sequenced and showed strong similarities with the globins of free-living and parasitic nematodes. Immunolocalization revealed the presence of this globin-like antigen in the body wall musculature and/or the cuticle of O. ostertagi adults. An enzyme-linked immunosorbent assay based on the purified OoAdGlb showed no differences in response between calves infected by O. ostertagi and/or Cooperia oncophora and the negative controls.

摘要

使用抗球蛋白兔血清Rb94进行的蛋白质印迹分析显示,在奥斯特他线虫成虫和第四期幼虫提取物中出现了一条主要的17 kDa条带。通过液相色谱法从全虫提取物中纯化出成虫期类球蛋白抗原(OoAdGlb)。在非还原条件下,该蛋白质的估计分子量为36 kDa,表明其为二聚体结构,包含2个非共价连接的17 kDa单体。对胰蛋白酶肽段进行了测序,结果显示其与自由生活和寄生线虫的球蛋白有很强的相似性。免疫定位显示,这种类球蛋白抗原存在于奥斯特他线虫成虫的体壁肌肉组织和/或角质层中。基于纯化的OoAdGlb的酶联免疫吸附测定表明,感染奥斯特他线虫和/或牛古柏线虫的犊牛与阴性对照之间的反应没有差异。

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