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牛坏死梭杆菌(AB)外膜 44.5kDa 蛋白的初步提取与鉴定

Preliminary Extraction and Identification of the 44.5 kDa Outer Membrane Proteins Isolated from Bovine Fusobacterium necrophorum (AB).

机构信息

College of Biology and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang, China ; State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Economic Animal and Plant Science, Chinese Academy of Agricultural Science, Changchun, China.

State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Economic Animal and Plant Science, Chinese Academy of Agricultural Science, Changchun, China.

出版信息

Indian J Microbiol. 2013 Dec;53(4):395-9. doi: 10.1007/s12088-013-0388-x. Epub 2013 Mar 16.

Abstract

Fusobacterium necrophorum (AB) in the pharynx, respiratory tract, female reproductive tract or urinary system is the causative agent of footrot and hepatic abscesses in animals and acute Lemierre's syndrome in humans. Current methods do not effectively protect animals and humans against F. necrophorum (AB). The outer membrane proteins (OMP) of F. necrophorum (AB) can be used as new material to protect against the diseases induced by F. necrophorum (AB). The aim of this study was to extract OMP and examine the immunogenic response of OMP. The preliminary extraction of OMP of F. necrophorum (AB) was identified by SDS-PAGE and stained by Coomassie Brilliant Blue R-250 (CB B R-250) and silver staining methods. The results showed that only a major band of 44.5 kDa was observed when staining the gel using CB B R-250. This band represented the target protein. In contrast, many small bands were observed by the silver staining method. The OMP also exhibited immune biological activities according to western blot analysis. The brightest band among the multi-banding observed was the OMP. Thus, the OMP was obtained and had immunogenic activity. The results provide a new direction to protect animals and humans against F. necrophorum (AB) in the clinical setting.

摘要

坏死梭杆菌(AB)存在于咽、呼吸道、生殖道或泌尿系统中,是导致动物腐蹄病和肝脓肿以及人类急性莱姆病的病原体。目前的方法不能有效地保护动物和人类免受坏死梭杆菌(AB)的侵害。坏死梭杆菌(AB)的外膜蛋白(OMP)可以作为一种新的材料,用于预防由坏死梭杆菌(AB)引起的疾病。本研究旨在提取 OMP 并检测 OMP 的免疫原性反应。采用 SDS-PAGE 对坏死梭杆菌(AB)的 OMP 进行初步提取,并采用考马斯亮蓝 R-250(CB B R-250)和银染法进行染色。结果表明,使用 CB B R-250 染色凝胶时仅观察到 44.5 kDa 的主要条带,该条带代表目标蛋白。相比之下,银染法观察到许多小条带。Western blot 分析表明 OMP 还具有免疫生物学活性。观察到的多带中最亮的条带是 OMP。因此,获得了具有免疫原性的 OMP。这些结果为临床预防动物和人类感染坏死梭杆菌(AB)提供了新的方向。

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