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革兰氏阴性菌细胞表面蛋白的免疫沉淀

Immunoprecipitation of Cell Surface Proteins from Gram-negative Bacteria.

作者信息

Cunha Carlos Eduardo Pouey, Newcombe Jane, Dellagostin Odir Antonio, McFadden Johnjoe

机构信息

School of Biosciences and Medicine, Faculty of Health and Medical Sciences, University of Surrey,Guildford, Surrey GU2 7XH, UK.

Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, Brasil.

出版信息

Bio Protoc. 2017 May 5;7(9):e2250. doi: 10.21769/BioProtoc.2250.

Abstract

The meningococcus () remains an important threat to human healthworldwide. This Gram-negative bacterium causes elevated disabilities and mortality in infectedindividuals. Despite several available vaccines, currently there is no universal vaccine against allcirculating meningococcal strains ( Vogel , 2013 ). Herein, we describe a new protocol that iscapable of identifying only cell surface exposed proteins that play a role in immunity, providing thisresearch field with a more straightforward approach to identify novel vaccine targets. Even though is used as a model in the protocol herein described, this protocol can be used for anyGram-negative bacteria provided modifications and optimizations are carried out to adapt it to differentbacterial and disease characteristics (, membrane fragility, growth methods, serum antibody levels,).

摘要

脑膜炎双球菌()仍然是全球人类健康的一个重要威胁。这种革兰氏阴性细菌会导致受感染个体的残疾率和死亡率上升。尽管有几种可用的疫苗,但目前尚无针对所有流行的脑膜炎球菌菌株的通用疫苗(Vogel,2013年)。在此,我们描述了一种新方案,该方案能够仅识别在免疫中起作用的细胞表面暴露蛋白,为该研究领域提供了一种更直接的方法来识别新型疫苗靶点。尽管本文所述方案中使用作为模型,但只要进行修改和优化以使其适应不同的细菌和疾病特征(如膜脆性、生长方法、血清抗体水平等),该方案可用于任何革兰氏阴性细菌。

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