【摘要】 根据禽巴氏杆菌5:A C48-1株OmpH全长基因序列设计一对特异性引物,经PCR扩增获得成熟外膜蛋白H基因(OmpmH),将OmpmH片段非定向插入原核表达载体pGEX-6p-1中,构建重组表达质粒pGEX-OmpmH。转化大肠杆菌BL21(DE3),在IPTG诱导下表达融合蛋白GST-OmpmH。SDS-PAGE结果显示,GST-OmpmH约为63 Ku,与预期的大小一致。Western blot检测结果表明,GST-OmpmH能与C48-1外膜蛋白免疫血清发生特异性反应,证明C48-1 ompmH原核融合表达成功。其可溶性蛋白经亲和层析纯化,得到了纯度较高的目的蛋白OmpmH。为进一步研究禽巴氏杆菌C48-1 OmpmH的免疫原性奠定了基础。更多还原

【Abstract】 According to C48-1 ompH nucleotide sequence,a pair of primers were designed,The mature outer membrane protein(ompmH) gene was amplified by PCR.It was cloned into the prokaryote expression vector pGEX-6p-1 and obtained recombinant plasmid pGEX-ompmH.pGEX-ompmH could express with fused protein GST-ompmH in E.coli BL21(DE3) via IPTG induction.The expressed product was identified by SDS-PAGE and Western-blotting.The resltuts showed that fusion protein GST-OmpmH was about 63 Ku in size and it could react with antisera of C48-1 outer membrane protein.After purification with affinity column and digested with PreScission Protease,OmpmH protein were obtained.These results lay the foundation for research on development of immuni-genicityofC48-1 OmpmH更多还原