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Outer Membrane Proteome Analysis of Indian Strain of Pasteurella multocida Serotype B:2 by MALDI-TOF/MS Analysis

机译:MALDI-TOF / MS分析印度多杀性巴氏杆菌B:2型印度菌株的外膜蛋白质组

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摘要

Identification of outer membrane proteins (OMPs) is important to understand the bacteria structure and function, host-pathogen interaction, development of novel vaccine candidates, and diagnostic antigens. But till now the key antigens of P. multocida B:2 isolate causing haemorrhagic septicaemia (HS) in animals are not clearly defined. In this study, P52 strain of P. multocida serotype B:2 was grown in vitro under iron-rich and iron-limited condition. The OMPs were extracted by sarkosyl method followed by SDS-PAGE and the proteins were identified by MALDI-TOF/MS analysis. In total, 22 proteins were identified, of which 7 were observed exclusively under iron-limited condition. Most of the high molecular weight proteins (TbpA, HgbA, HgbB, HasR, IroA, and HemR) identified in this study were involved in iron acquisition. Some hypothetical proteins (HP-KCU-10206, HP and AAUPMB 08244, HP AAUPMB 21592, HP AAUPMB 19766, AAUPMB 11295) were observed for the first time in this study which could be unique to serotype B:2. Further functional in vivo study of the proteins identified are required to explore the utility of these proteins in developing diagnostics and vaccine against HS.
机译:鉴定外膜蛋白(OMP)对于了解细菌的结构和功能,宿主与病原体的相互作用,新型疫苗候选物的开发以及诊断抗原非常重要。但是到目前为止,尚不清楚对导致动物出血性败血病(HS)的多杀青霉菌B:2分离株的关键抗原的定义。在这项研究中,在富含铁和有限铁的条件下,在体外培养了P. multocida血清型B:2的P52菌株。通过Sarkosyl方法提取OMP,然后进行SDS-PAGE提取,并通过MALDI-TOF / MS分析鉴定蛋白质。总共鉴定出22种蛋白质,其中仅在铁限制条件下观察到7种蛋白质。这项研究中鉴定的大多数高分子量蛋白质(TbpA,HgbA,HgbB,HasR,IroA和HemR)都与铁的摄取有关。在本研究中首次观察到某些假设蛋白(HP-KCU-10206,HP和AAUPMB 08244,HP AAUPMB 21592,HP AAUPMB 19766,AAUPMB 11295),这些蛋白可能是血清型B:2特有的。需要进一步对鉴定出的蛋白质进行功能性体内研究,以探索这些蛋白质在开发针对HS的诊断方法和疫苗中的实用性。

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