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脑膜炎奈瑟菌PorA外膜的计算机模拟研究:其表达及免疫原性特性

In silico studies of outer membrane of Neisseria meningitidis por a: its expression and immunogenic properties.

作者信息

Behrouzi Ava, Bouzari Saeid, Siadat Seyed Davar, Irani Shiva

机构信息

Department of Biology, School of Basic Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.

Department of Biology, Pasteur Institute of Iran, Tehran, Iran.

出版信息

Int J Mol Cell Med. 2014 Summer;3(3):166-75.

Abstract

Neisseria meningitidis is a major causative agent of bacterial septicemia and meningitis in humans. Currently, there are no vaccines to prevent disease caused by strains of N.meningitidis serogroup B. The Class 1 Outer Membrane Protein (OMP) has been named porA which is a cation selective transmembrane protein of 45 KDa that forms trimeric pore in the meningococcal outer membrane. PorA from serogroup B N. meningitidis was cloned into prokaryotic expression vector pBAD-gIIIA. Recombinant protein was expressed with arabinose and affinity purified by Ni-NTA agarose, SDS-PAGE and western blotting were performed for protein determination and verification. BALB/c mice were immunized subcutaneously with purified rPorA together with alum adjuvant. Serum antibody responses to serogroups B N.meningitidis were determined by ELISA. Serum IgG response significantly increased in the group immunized with rPorA together with alum adjuvant in comparison with control groups. These results suggest that rPorA can be a potential vaccine candidate for serogroup B N.meningitidis.

摘要

脑膜炎奈瑟菌是人类细菌性败血症和脑膜炎的主要病原体。目前,尚无预防B群脑膜炎奈瑟菌菌株所致疾病的疫苗。1类外膜蛋白(OMP)被命名为PorA,它是一种45 kDa的阳离子选择性跨膜蛋白,在脑膜炎球菌外膜中形成三聚体孔道。将B群脑膜炎奈瑟菌的PorA克隆到原核表达载体pBAD-gIIIA中。用阿拉伯糖表达重组蛋白,并通过Ni-NTA琼脂糖进行亲和纯化,进行SDS-PAGE和蛋白质印迹以进行蛋白质测定和验证。用纯化的rPorA与明矾佐剂一起皮下免疫BALB/c小鼠。通过ELISA测定对B群脑膜炎奈瑟菌的血清抗体反应。与对照组相比,用rPorA与明矾佐剂免疫的组中血清IgG反应显著增加。这些结果表明,rPorA可能是B群脑膜炎奈瑟菌的潜在疫苗候选物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/440b/4170490/5792a59a1adf/ijcm-3-166-g001.jpg

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