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犬钩口线虫重组副肌球蛋白的体外表达

In vitro expression of a recombinant paramyosin of Ancylostoma caninum.

作者信息

Kohlmetz C, Epe C, Schnieder T

机构信息

Hannover School of Veterinary Medicine, Institute of Parasitology, Germany.

出版信息

Int J Parasitol. 1998 Aug;28(8):1229-33. doi: 10.1016/s0020-7519(98)00106-4.

Abstract

The objective of this study was to characterise a recombinant antigen of Ancylostoma caninum that had been identified by immunoscreening with selected antisera described elsewhere. In vitro expression of clone 341 produced a protein with an apparent molecular mass of approximately 34 kDa which was recognised in Western blots by antisera against whole worms and antisera against esophagi from adult worms, but not by sera from experimentally infected dogs or rabbits. DNA sequencing showed a cDNA of 1176 bp coding for a 34-kDa protein, similar to the size identified in the immunoblot. DNA database comparison revealed an 80-82% homology with the Caenorhabditis elegans unc-15 gene coding for paramyosin. The deduced aa sequence of clone 341 showed 95% homology with the paramyosin aa sequence of C. elegans. Affinity purified antibodies against the recombinant protein recognise a protein with an apparent molecular mass of 97 kDa of A. caninum muscle tissue fraction which is in accordance with the molecular mass of paramyosin from Schistosoma mansoni and Schistosoma japonicum.

摘要

本研究的目的是对犬钩口线虫的一种重组抗原进行特性鉴定,该抗原是通过用其他地方描述的选定抗血清进行免疫筛选而鉴定出来的。克隆341的体外表达产生了一种表观分子量约为34 kDa的蛋白质,在蛋白质印迹中,该蛋白质可被抗全虫抗血清和抗成虫食管抗血清识别,但不能被实验感染犬或兔的血清识别。DNA测序显示一个1176 bp的cDNA编码一种34 kDa的蛋白质,与免疫印迹中鉴定的大小相似。DNA数据库比较显示,其与编码副肌球蛋白的秀丽隐杆线虫unc-15基因有80-82%的同源性。克隆341推导的氨基酸序列与秀丽隐杆线虫的副肌球蛋白氨基酸序列有95%的同源性。针对该重组蛋白的亲和纯化抗体识别犬钩口线虫肌肉组织部分中一种表观分子量为97 kDa的蛋白质,这与曼氏血吸虫和日本血吸虫副肌球蛋白的分子量一致。

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