Cloning, expression and characterization of a cell wall surface protein, 6-phosphogluconate dehydrogenase, of Haemophilus parasuis
Cloning, expression and characterization of a cell wall surface protein, 6-phosphogluconate dehydrogenase, of Haemophilus parasuis
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DOI：
10.1016/j.rvsc.2011.07.006
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发表时间：
2012-08-01
影响因子：
2.4
通讯作者：
Bei, Weicheng
中科院分区：
文献类型：
--
作者：
Fu, Shulin;Yuan, Fangyan;Bei, Weicheng
副猪嗜血杆菌（Haemophilus parasuis，H.副猪链球菌（parasuis）是引起格拉瑟氏病的猪病原体。为了解H. god基因编码一种细胞表面蛋白6-磷酸脱氢酶（6-PGD）。在大肠杆菌BL 21中诱导表达具有六组氨酰N-末端的副猪嗜血杆菌，以允许其纯化。用r6 PGD特异性抗血清进行的Western印迹分析表明，6PGD蛋白位于H.副猪。对H.并进一步研究了其在小鼠体内的免疫原性。粘附试验用H.用（His）（6）6PGD预孵育的副猪和猪肺泡上皮细胞（SJPLC）和未孵育的SJPLC显示H.与未孵育的SJPLC相比，（His）（6）6PGD预孵育的SJPLC中的副猪嗜血杆菌。此外，r6 PGD蛋白通过SJPLC诱导IL-8和IL-6的产生。此外，用r6 PGD蛋白免疫可以在腹膜内施用5 × LD 50剂量的H. parasuis SH 0165的6PGD表达，并诱导了良好的保护性免疫应答，证明了6PGD在细菌致病过程中的重要性。鉴定和表征H. parasuis 6PGD在粘附和免疫原性方面的研究将使我们能够使用该蛋白开发新的抗微生物疗法和/或疫苗。(C)2011爱思唯尔有限公司版权所有。
Haemophilus parasuis (H. parasuis) is a swine pathogen responsible for the Glasser's disease. In order to understand the pathogenesis of the H. parasuis infection, the god gene encoding a cell surface protein, 6-phosphogluconate-dehydrogenase (6PGD) of H. parasuis was inducibly expressed in Escherichia coli BL21 with a hexahistidyl N-terminus to permit its purification. Western blotting using the r6PGD-specific antiserum showed that the 6PGD protein is on the cell surface of H. parasuis. The characterization of 6PGD in H. parasuis pathogenesis involved as an adhesion and its immunogenicity in mice was further investigated. The adherence assay with H. parasuis and swine alveolar epithelial cells (SJPLC) pre-incubated with (His)(6)6PGD and non-incubated SJPLC showed a noticeable reduction in the adhesion of H. parasuis in the (His)(6)6PGD pre-incubated SJPLC compared to the non-incubated SJPLC. Further, the r6PGD protein induces the production of IL-8 and IL-6 by SJPLC. Furthermore, immunization with the r6PGD protein can provide the protective efficacy by 75% following intraperitoneal administration of a 5 x LD50 dose of H. parasuis SH0165, and elicited a good protective immune response, which demonstrated the importance of 6PGD to bacterial pathogenesis. Identification and characterization of the role of H. parasuis 6PGD in adhesion and immunogenicity will allow us to use this protein to develop new antimicrobial therapies and/or vaccines. (C) 2011 Elsevier Ltd. All rights reserved.