禽巴氏杆菌C48-1株成熟外膜蛋白H基因的原核融合表达

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63Ku，与预期的大小一致。Westernblot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C州OmpmH的免疫原性奠定了基础。

Fusion expression of mature outer membrane protein H gene of avian pasteurella multocida C48-1 in Escherichia coli

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