K88菌毛基因的克隆及ST表位序列在菌毛基因上的融合  

Cloning of K88 Pili Gene and Fusion of ST Epitope Coding Sequence in Pili Gene

作　　者：胡建军[1] 陈创夫[2] 祝建波[1] 

机构地区：[1]石河子大学农业生物技术重点实验室,新疆石河子832003 [2]石河子大学动物科技学院,新疆石河子832003

出　　处：《安徽农业科学》2008年第21期8949-8951,共3页Journal of Anhui Agricultural Sciences

摘　　要：[目的]为了建立用于细胞表面展示的载体系统。[方法]利用PCR技术,以大肠杆菌质粒为模板扩增K88菌毛操纵子的结构基因faeC^faeH,并克隆到pBR322质粒载体中。合成一段替换序列,在菌毛抗原基因的超变区引入酶切位点ApaI和NcoI,合成耐热肠度素STII表位编码序列,并引入菌毛抗原基因的超变区。[结果]经PCR鉴定和限制性内切酶酶切证明,重组质粒pBR-fae插入片断大小为6.6kb,与预期相符。核苷酸序列分析证明,所得faeC^faeH序列正确。PCR筛选和测序验证证明,构建了K88菌毛-耐热肠度素STII的融合基因。[结论]试验成功获得了重组质粒pBR-fae-ST。[Objective] The objective was to meet the requirements of establishing the cell surface display system.[Method]PCR was used to amplify the structural gene faeC-faeH of K88 pili operon,and then the structural gene was cloned into the vector pBR322.The sequence containing restriction sites ApaI and NcoI was synthesized and fused in the supper variable region of K88 pili antigen gene.And the ST epitope coding sequence was synthesized and fused in the restriction sites ApaI and NcoI.[Result] PCR and restriction enzyme digestion demonstrated that the inserted fragment was 6.6 kb in recombinant plasmid pBR-fae.Nucleotide sequence analysis demonstrated the correctness of faeC-faeH.PCR screening and sequence analysis verified that the K88 pili-STII fusion gene was constructed.[Conclusion] The recombinant plasmid pBR-fae-ST was successfully constructed in this experiment.

关 键 词：肠毒素大肠杆菌 K88菌毛 ST表位表位 

分 类 号：S188[农业科学—农业基础科学]