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猪链球菌2型细胞壁表面蛋白6-磷酸葡萄糖酸脱氢酶的克隆、表达及特性分析

Cloning, expression and characterization of a cell wall surface protein, 6-phosphogluconate-dehydrogenase, of Streptococcus suis serotype 2.

作者信息

Tan Chen, Fu Shulin, Liu Manli, Jin Meilin, Liu Jinlin, Bei Weicheng, Chen Huanchun

机构信息

Division of Animal Pathogens, State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, PR China.

出版信息

Vet Microbiol. 2008 Aug 25;130(3-4):363-70. doi: 10.1016/j.vetmic.2008.02.025. Epub 2008 Mar 6.

Abstract

Streptococcus suis type 2 is a pathogen responsible for diverse diseases in both pigs and humans. In order to understand the pathogenesis of the S. suis type 2 infection, the gene encoding a cell surface protein, 6-phosphogluconate-dehydrogenase (6PGD) of S. suis type 2 was cloned and sequenced, and recombinant 6PGD protein (r6PGD) was produced in a prokaryotic expression system. Sequence analysis of the cloned 6 pdg gene showed 82% similarity with Streptococcus pneumoniae 6 pdg at the nucleic acid level. Western blotting using r6PGD-specific antiserum confirmed the cell surface location of the 6PGD protein of S. suis type 2. The role of 6PGD in S. suis type 2 pathogenesis as an adhesin and its immunogenicity in mice was further investigated. The results showed that the recombinant protein interfered with the adhesion of S. suis type 2 to Hep2 and HeLa cells by 72% and 66%, respectively. Immunization of CD-1 mice with r6PGD increased the protective efficacy by 80% following intraperitoneal administration of a lethal dose of S. suis type 2. Immunization of CD-1 mice with r6PGD elicited a significant protective immune response, which demonstrated the importance of 6PGD to bacterial pathogenesis. Identification and characterization of the role of S. suis type 2 6PGD in adhesion and immunogenicity will allow us to use this protein to develop new antimicrobial therapies and/or vaccines.

摘要

猪链球菌2型是一种可导致猪和人类多种疾病的病原体。为了解猪链球菌2型感染的发病机制,克隆并测序了编码猪链球菌2型细胞表面蛋白6-磷酸葡萄糖酸脱氢酶(6PGD)的基因,并在原核表达系统中生产了重组6PGD蛋白(r6PGD)。对克隆的6pdg基因的序列分析表明,其在核酸水平上与肺炎链球菌6pdg的相似性为82%。使用r6PGD特异性抗血清进行的蛋白质印迹证实了猪链球菌2型6PGD蛋白在细胞表面的定位。进一步研究了6PGD在猪链球菌2型发病机制中作为黏附素的作用及其在小鼠中的免疫原性。结果表明,重组蛋白分别使猪链球菌2型对Hep2和HeLa细胞的黏附减少了72%和66%。用r6PGD免疫CD-1小鼠,在腹腔注射致死剂量的猪链球菌2型后,保护效力提高了80%。用r6PGD免疫CD-1小鼠引发了显著的保护性免疫反应,这证明了6PGD对细菌发病机制的重要性。鉴定和表征猪链球菌2型6PGD在黏附和免疫原性方面的作用,将使我们能够利用这种蛋白质开发新的抗菌疗法和/或疫苗。

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