犬布氏杆菌外膜蛋白Omp31基因的原核表达

克隆了犬布氏杆菌外膜蛋白Omp31基因并构建原核表达系统,并对表达产物进行了初步的血清学鉴定。利用PCR技术扩增犬布氏杆菌RM6/66参考株Omp31基因,然后将其克隆到pGEMT-easy载体上进行测序。测序正确后,将该基因插入到pET-32a载体中构建原核表达载体,转化大肠杆菌BL21感受态细胞,诱导表达融合蛋白,Western blot分析融合蛋白的免疫反应性。结果构建了犬布氏杆菌Omp31基因的原核表达载体pET-Omp31,并且在大肠杆菌中成功表达融合蛋白,经Western Blot鉴定该蛋白能被犬布氏杆菌阳性血清所识别。犬布氏杆菌外膜蛋白Omp31的表达成功,为犬布氏杆菌病血清学诊断方法的建立提供了基础资料。

Prokaryotic expression of outer membrane protein Omp31 gene of Brucella canis

The gene of outer membrane protein Omp31 was amplified by PCR from Brucella canis strain RM6/66.After cloning and sequencing,the gene was integrated into pET-32a vector.Recombinant plasmid pET-Omp31 was transformed into E.coli BL21(DE3) competent cells and induced to express Omp31 by IPTG.The fusion protein as expected by SDS-PAGE was produced and Western blot result showed that the expression product could specifically react with the antiserum to B.canis.This study will be helpful for establishing new serum diagnostic methods of B.canis infection.