空肠弯曲菌peb1A基因的克隆表达及免疫原性分析  

Cloning,Expression and Immunogenicity Analysis of Campylobacter jejuni peb1A Gene

作　　者：祝长青[1,2] 唐泰山[1,2] 王婷[3] 蒋原[1] 姚火春[2] 张常印[1] 薛峰[1] 栾军[1] 

机构地区：[1]江苏出入境检验检疫局,江苏南京210001 [2]南京农业大学动物医学院,江苏南京210095 [3]河南科技大学动物科技学院,河南洛阳471003

出　　处：《动物医学进展》2011年第9期1-4,共4页Progress In Veterinary Medicine

基　　金：国家质检总局科研项目(2008IK145)

摘　　要：根据空肠弯曲菌基因序列设计引物,以菌株ATCC 29428为模板扩增出peb1A基因片段,定向克隆至pMD18-T载体中,并对克隆片段测序,比较所测序列与不同来源弯曲菌的同源性;用软件改造并合成peb1A基因78 bp^780 bp区间片段,PCR扩增后,构建出表达载体pET-28a(+)-peb1Ag,转化至E.coliBL21(DE3)后诱导表达,纯化后获得重组蛋白PEB1;将其进行Western blot分析、小鼠免疫试验。结果表明,克隆的peb1A基因序列与报道的NCTC 11168核苷酸同源性为98.1%,改造表达后的重组蛋白PEB1具有良好的抗原性和免疫原性,免疫小鼠后血清效价达1∶12 800,为空肠弯曲菌保护性抗原的研究和单克隆抗体的筛选奠定了基础。Based on the C.jejuni genes in GenBank,the peb1A genes was amplified by PCR from C.jejuni genomic DNA.The PCR fragments were cloned into the pMD18-T vector and sequenced.The sequences were compared with the peb1A genes of different kinds of C.jejuni strains to analyze their homology and diversity degree.The DNA fragment of peb1Ag gene of C.jejuni was optimized and synthesized in accordance with the E.coli ＇s codon preference,it was inserted to the expression vector pET-28a（＋）.The recombinant expression plasmid pET-28a（＋）-peb1Ag was transformed into E.coli BL21（DE3）,and the recombinant protein PEB1 was expressed,purified,and detected by Western blot and the mouse immunologic test.The results showed the recombinant protein PEB1 possessed the good immunogenicity.

关 键 词：空肠弯曲菌 peb1A 原核表达 免疫原性 

分 类 号：S852.613[农业科学—基础兽医学]