布氏菌外膜蛋白bp26的表达和鉴定

EXPRESSION AND IDENFICATION OF BRUCELLA OUTER MEMBRANE PROTEIN bp26

[目的]研究布鲁氏菌外膜蛋白bp26基因的克隆、表达与测序,并对其进行初步的血清学鉴定。[方法]从布鲁氏菌基因组中获得bp26基因连接入PMD18-T克隆质粒并测序,目的基因与融合表达载体PGEX-4T-1连接,构建重组质粒PGEX-4T-1/bp26,在大肠杆菌中表达该蛋白。用western-blot鉴定重组表达的GST-bp26蛋白。[结果]成功构建了PGEX-4T-1/bp26原核表达载体,并在大肠杆菌中成功表达了bp26基因,布鲁氏菌免疫动物血清能特异性识别所表达的蛋白。[结论]布鲁氏菌外膜蛋白bp26基因的成功表达,它可以与布鲁氏菌免疫血清产生特异性结合反应,为之后的抗原性以及免疫原性研究打下良好的物质基础。

[Objective] To study the cloning sequencing and expression of Brucella outer membrane were protein bp26, and then detect its reactivity in serum from Brucella immuned rabbit. [ Methods] The bp26 gene were got from Brucella genome and cloned into PMD18-T. After sequencing, the bp26 gene were cloned into vector PGEX-4T-1 and expressed in E coli, then detected by western-blot. [Results] According to the SDS-PAGE and western-blot analysis, the recombinant plasmid PGEX-4T-1/bp26 could be expressed successfully in E eoli and the fusion protein could react with serum from Brueella immuned rabbit. [ Conclusion ] The fusion protein GST-bp26 can react with serum from Brucella immuned rabbit, which provides good foundation for the study of lab diagnosis and preparation of submit vaccine.