Immunogenicity and immunoprotection of recombinant PEB1 in Campylobacter-jejuni-infected mice  

作　　者：Lian-Feng Du Zhen-Jiang Li Xian-Ying Tang Jun-Qiong Huang Wan-Bang Sun 

机构地区：[1]Department of Immunology, ZhuhaiCampus of Zunyi Medical College, Zhuhai 519041, GuangdongProvince, China

出　　处：《World Journal of Gastroenterology》2008年第40期6244-6248,共5页世界胃肠病学杂志（英文版）

基　　金：Grants from the Governor Foundation for Excellent Talents of Guizhou Province,No.200607

摘　　要：AIM： To construct a prokaryotic expression vector carrying Campylobacterjejuni peblA gene and express it in Escherichia coli. Immunoreactivity and antigenicity of rPEB1 were evaluated. The ability of rPEB1 to induce antibody responses and protective efficacy was identified. METHODS： peblA gene was amplified by PCR, target gene and prokaryotic expression ptasmid pET28a （＋） was digested with BamHI and XhoI, respectively. DNA was ligated with T4 DNA ligase to construct recombinant plasmid pET28a（＋）-peblA. The rPEB1 was expressed in E. coli BL21 （DE3） and identified by SDS-PAGE. BALB/c mice were immunized with rPEBI. ELISA was used to detect the specific antibody titer and MTT method was used to measure the stimulation index of spleen lymphocyte transformation. RESULTS： The recombinant plasmid pET28a （＋）-peblA was correctly constructed. The expression output of PEB1 protein in pET28a （＋）-peblA system was approximately 33% of total proteins in E. coil The specific IgG antibody was detected in serum of BALB/c mice immunized with rPEB1 protein. Effective immunological protection with a lower sickness incidence and mortality was seen in the mice suffering from massive C. jejuni infection. CONCLUSION： rPEB1 protein is a valuable candidate for C. jejuni subunit vaccine.AIM: To construct a prokaryotic expression vector carrying Campylobacter jejuni peb1A gene and express it in Escherichia coli.Immunoreactivity and antigenicity of rPEB1 were evaluated.The ability of rPEB1 to induce antibody responses and protective efficacy was identified.METHODS: peb1A gene was amplified by PCR,target gene and prokaryotic expression plasmid pET28a (+) was digested with BamHI and XhoI,respectively.DNA was ligated with T4 DNA ligase to construct recombinant plasmid pET28a(+)-peb1A.The rPEB1 was expressed in E.coli BL21 (DE3) and identified by SDS-PAGE.BALB/c mice were immunized with rPEB1.ELISA was used to detect the specific antibody titer and MTT method was used to measure the stimulation index of spleen lymphocyte transformation.RESULTS: The recombinant plasmid pET28a (+)-peb1A was correctly constructed.The expression output of PEB1 protein in pET28a (+)-peb1A system was approximately 33% of total proteins in E.coli.The specific IgG antibody was detected in serum of BALB/c mice immunized with rPEB1 protein.Effective immunological protection with a lower sickness incidence and mortality was seen in the mice suffering from massive C.jejuni infection.CONCLUSION: rPEB1 protein is a valuable candidate for C.jejuni subunit vaccine.

关 键 词：Campy/obacterjejuni Prokaryotic expression IMMUNOGENICITY 

分 类 号：R392[医药卫生—免疫学]