【摘要】 目的研究布鲁氏菌外膜蛋白OMP31的克隆、测序和表达,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OMP31基因连接入PMD-18T克隆质粒并测序,克隆入融合表达载体PGEX-4T-1,构建重组质粒PGEX-4T-1/OMP31,在大肠杆菌中将该蛋白表达。用Western-blot分析重组表达的GST-OMP31的免疫学特性。结果成功构建了PGEX-4T-1/OMP31原核表达载体并在大肠杆菌中表达了OMP31基因,布鲁氏菌免疫动物血清能特异性识别所表达的蛋白。结论成功表达了布鲁氏菌外膜蛋白OMP31基因,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基础。更多还原

【Abstract】 Objective To clone and express the brucella outer membrane protein omp31 and then detect its reactivity to the Brucella immunized rabbit serum. Method The OMP31 gene of Brucella was amplified from genomic DNA and cloned into PMD-18T vector. It was then cloned into vector pGEX-4T-1 and expressed in E.coli. The recombinant protein was detected by Western-blot. Result According to the SDS-PAGE and Western-blot analysis, the recombinant plasmid pGEX-4T-1/OMP31 could be expressed successfully in E.coli and the fusion protein could react with serum from Brucella immunized rabbit. Conclusion The fusion protein GST-OMP31 could react with serum from Brucella immunized rabbit. This result can facilitate the characterization of the antigen and preparing subunit vaccine.更多还原