脑膜炎球菌外膜蛋白NhhA的表达及免疫效果评价  

Expression of Meningococcal Outer Membrane Protein NhhA and Its Immunological Effect

作　　者：王慧敏[1] 高云英[1] 田飞鹏[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨陵712100

出　　处：《动物医学进展》2010年第12期53-57,共5页Progress In Veterinary Medicine

摘　　要：NhhA是在对脑膜炎球菌MC58株进行全基因组测序过程中发现的膜表面蛋白质,属于三聚体自体转运黏附素家族,介导细菌黏附于宿主鼻黏膜上皮细胞。为了对NhhA在小鼠体内诱生体液免疫效果进行评价,将截去信号肽编码序列的目的基因克隆到原核表达载体pET-21b(+)上,3′端与6×His标签融合表达,用Ni柱螯合层析法纯化目的蛋白,并用脑膜炎球菌A、B、C、W135、Y群的混合免疫血清对重组目的蛋白进行免疫印迹鉴定,结果均呈阳性反应。以Al(OH)3和CpG作为NhhA的佐剂免疫Balb/c小鼠,二次免疫后采血进行ELISA检测,其血清抗体滴度高达1∶100 000。用脑膜炎球菌裂解物鉴定免疫血清的特异性,结果显示为阳性。NhhA（GNA992）,is a surface protein that was identified during the Neisseria meningitidis group B strain MC58 genome-sequencing project.It is included in the family of trimeric autotransporter adhesins,and immediate bacterial adhesion to and penetration into human epithelial cells.To evaluate the immunological effect of NhhA in mice,the gene was cloned and truncated into the pET-21b（＋） expression vector,and the carboxyl-terminal ends fused to 6xHis tag.Protein was purified by Ni＋ metal chelate chromatography,and the protein showed positive activity in Western blot using meningococcal anti-serum as detecting antibody.Al（OH）3 and CpG-ODN（CpG oligodeoxynucleotides） were used as adjuvants of NhhA to immunize Balb/c mice.The sera antibody titers were detected with ELISA after second immunization in mice.The results indicated that the antibody titers induced by NhhA were 1∶100 000,the serum specificity was detected using Western blot and the result was positive.

关 键 词：脑膜炎球菌 NhhA 纯化 表达 免疫 

分 类 号：Q786[生物学—分子生物学]