【摘要】 将Omp L17基因克隆于p GEX- 1λT载体,在大肠杆菌JM10 9中表达分子量约为5 4 KDa的GST-Omp L17融合蛋白,凝血酶切割后得到了大小约2 8KDa的Omp L17蛋白。用纯化的Omp L17蛋白免疫大白兔,制备了高滴度的特异性抗体(1∶4 896 )。本研究获得了纯化的Omp L17及其特异性抗体,为该外膜蛋白致病作用及免疫保护力研究奠定基础。更多还原

【Abstract】 This study was conducted to potentiate the expression of outer membrane protein OmpL17 of the strong virulent L. in terrogans serovar Lai and investigate its immunogenicity in rabbits. The OmpL17 was cloned into prokaryotic expression vector pGEX-1λT. The recombination expr e ssion plasmid pGEX-OmpL17 was transformed into E.Coli JM109. The GST fused pro t ein GST-OmpL17 was expressed after induction by IPTG, then GST-tag was by th rombin and purified using Bulk GST purification Modules. SDS-PAGE and Western b l otting analysis indicated that the molecular weight of GST-OmpL17 and OmpL17 wa s about 54KDa and 28KDa respectively. The outer membrane protein OmpL17 was subcu taneously injected into rabbits and high titre anti-OmpL17 antibody was obtaine d (1∶4896) which could conjugate specifical with OmpL17. In conclusion, OmpL17 and specifical anti-OmpL17 antibody were obtained, which provided an experim ental basis for researching pathogenic effect and immunity functions of OmpL17.更多还原