布鲁氏菌omp28基因的克隆、表达及反应原性分析

本研究克隆了羊种布鲁氏菌16M株、羊种布鲁氏菌M28株、犬种布鲁氏菌、绵羊附睾种布鲁氏菌、牛种布鲁氏菌A19株、猪种布鲁氏菌S2株的omp28基因并对以上不同种菌株的omp28基因序列及编码的氨基酸序列进行了比对,结果显示不同种布鲁氏菌omp28基因之间仅6个碱基不同,而且只有2个氨基酸不同,亲水性分析结果显示两处氨基酸的差异对蛋白亲水性不造成影响.将羊种布鲁氏菌16M的omp28基因亚克隆到pET32a中表达,OMP28在低温下诱导以可溶性形式高效表达.Westem-blot结果显示OMP28反应原性良好,是布鲁氏菌病诊断抗原的可能选择.

Cloning and Expression of Brucella omp28 Gene and Analysis on Reactogenicity of Recombinant Brucella OMP28

In this study, omp28 genes of different brucella species including brucella melitensis 16M, brucella melitensis M28, brucella canis, brucella ovis, brucella abortus A19, brucella suis S2, were cloned and sequenced. The sequence alignment results of amino acid and nucleotide showed that 6 base pairs and 2 amino acids were different among omp28 genes in different brucella species, however had no influence to the hydrophily of OMP28. Omp28 gene of Brucella melitensis 16M were cloned into pET32a plasmid and expressed in vitro, OMP28 which was expressed in a soluable form at lower tempreture, was identified by western-blot, and the result showed that recombined protein obtained has a good reactogenicity and will be a good potential antigen for brucellosis diagnosis.