布氏杆菌L7/L12和OMP31基因克隆及融合表达  

Clone and Fusion Expression of L7/L12 and OMP31 Genes from Brucella

作　　者：李海涛[1] 苗利光[1] 张广雷[1] 刘艳环[1] 孙金霞[1] 王松[1] 王文昊[1] 

机构地区：[1]中国农业科学院特产研究所,吉林吉林132109

出　　处：《特产研究》2010年第4期4-8,共5页Special Wild Economic Animal and Plant Research

基　　金：国家科技支撑计划(2006BAD06B06)

摘　　要：本研究根据已经发表的布氏杆菌基因序列,选择种间序列极为保守的保护性抗原L7/L12、OMP31基因,依此进行PCR扩增,利用引物上设计好的限制性内切酶酶切位点进行基因重组,构建了PME290-SDLOmp高效表达载体,并转化绿脓杆菌(PAK/2pfs),通过培养得到了表达产物,对表达产物进行SDS-PAGE和Western Blot鉴定,结果符合预期。本项研究为进一步开展布氏杆菌亚单位疫苗的研制奠定了基础。In this study we carried out gene cloning and fusion expression according to the published gene sequence.We chose the protective antigens L7/L12 and OMP31 which were both extremely conserved sequences between species and amplify with PCR technology.We used the restriction sites designed on the primer carrying on gene recombination.We constructed the effective express vector PME290-SDLOmp.We transformed competence（PAK/2PFS）with constructed vector.After expansion of the bacteria we obtained the expression product.Subsequent characterization by the SDS-PAGE and Western Blot showed that we had the correct product.Our study had laid foundation for further develop of brucella subunit vaccine.

关 键 词：布氏杆菌 基因重组 表达载体构建 融合蛋白 

分 类 号：Q785[生物学—分子生物学]