In silico analysis and recombinant expression of BamA protein as a universal vaccine against Escherichia coli in mice
In silico analysis and recombinant expression of BamA protein as a universal vaccine against Escherichia coli in mice
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BamA 蛋白作为小鼠大肠杆菌通用疫苗的计算机分析和重组表达
DOI：
10.1007/s00253-016-7467-y
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发表时间：
2016-06-01
影响因子：
5
通讯作者：
Wang, Jianhua
中科院分区：
文献类型：
--
作者：
Guan, Qingfeng;Wang, Xiao;Wang, Jianhua
大肠杆菌病（Colibacillosis）是由致病性大肠杆菌（Escherichia coli）引起的一种世界性的动物和人类常见病，给养殖业造成巨大损失，每年造成数百万人死亡。因此，迫切需要开发针对大肠杆菌病的通用疫苗。在这项研究中，BamA蛋白的序列同源性，物理化学性质，过敏性预测和表位预测进行了分析。BamA蛋白（含286个氨基酸）在E.在UniProtKB数据库中检索到81.7%的同源性（Uniref entry A7 ZHR 7），其中同源性为94.82%的序列在93.4%以上。此外，BamA在沙门氏菌和志贺氏菌中高度保守，对小鼠和人无致敏性。BamA的抗原表位主要位于细胞周质和细胞外区。将BamA的Surf_Ag_VNR结构域（448-810 aa）表达、纯化，然后用于免疫小鼠。rBamA和E.对沙门氏菌和志贺氏菌的抑制率均在1：27，000以上。调理吞噬实验结果表明，rBamA血清可增强中性粒细胞对E.杆菌rBamA和PBS免疫小鼠的存活率分别为80%和20%。这些数据表明，BamA可以作为一个有前途的通用疫苗候选人的保护性亚单位疫苗的细菌感染的发展。该方案为有效控制病原性大肠杆菌提供了更多可行的技术线索和选择。大肠杆菌、沙门氏菌和志贺氏菌。
Colibacillosis, caused by pathogenic Escherichia coli, is a common disease in animals and human worldwide with extensive losses in breeding industry and with millions of people death annually. There is thus an urgent need for the development of universal vaccines against colibacillosis. In this study, the BamA protein was analyzed in silico for sequence homology, physicochemical properties, allergenic prediction, and epitopes prediction. The BamA protein (containing 286 amino acids) clusters in E. coli were retrieved in UniProtKB database, in which 81.7 % sequences were identical (Uniref entry A7ZHR7), and sequences with 94.82 % identity were above 93.4 %. Moreover, BamA was highly conserved among Salmonella and Shigella and has no allergenicity to mice and human. The epitopes of BamA were located principally in periplasm and extracellular domain. Surf_Ag_VNR domain (at position 448-810 aa) of BamA was expressed, purified, and then used for immunization of mice. Titers of the rBamA sera were 1:736,000 and 1:152,000 against rBamA and E. coli and over 1:27,000 against Salmonella and Shigella. Opsonophagocytosis result revealed that the rBamA sera strengthened the phagocytic activity of neutrophils against E. coli. The survival rate of mice vaccinated with rBamA and PBS was 80 and 20 %, respectively. These data indicated that BamA could serve as a promising universal vaccine candidate for the development of a protective subunit vaccine against bacterial infection. Thus, the above protocol would provide more feasible technical clues and choices for available control of pathogenic E. coli, Salmonella, and Shigella.