新疆绵羊种布鲁氏菌外膜蛋白OMP36的表达蛋白OMP2b基因的克隆与表达

以表达新疆绵羊种布鲁氏菌外膜抗原蛋白OMP36的表达蛋白OMP2b,探索其作为诊断抗原和分子疫苗的可能性。采用PCR扩增技术,从新疆绵羊种布鲁氏菌基因组DNA中扩增出OMP2b基因片段,将该片段克隆于原核表达载体PE-28a(+),构建成重组质粒,IPTG诱导表达,SDS-PAGE检测有无蛋白的表达。结果表明,获得长约1 083bp的PCR片段,序列分析结果与已知绵羊种OMP2b同源性达89.72%;SDS-PAGE检测表达产物,在相对分子量39 ku获得了新疆绵羊种布鲁氏菌外膜蛋白OMP36的表达蛋白OMP2b基因片段,并在大肠杆菌中实现了表达。

Cloning and Expressing of out Membrane Protein OMP2b from Brucella ovis in Xinjiang

Objective: to express OMP2b antigen protein of OMP36 from out membrane protein of Brucella ovis in XinJiang and study the probability for it as molecular bacterin and diagnose antigen.Methods : OMP2b protein gene fragment was amplified from genomic DNA of Brucella ovis in XinJiang through PCR.The fragment was identified and cloned into prokaryotic expression vector PET-28a(+).Then IPTG was used to induce expression of OMP2b antigen protein.Result:(1)An about 1 089bp length PCR product was obtained and the sequence was the same as OMP2b of B.ovis sequence reported before.(2)An expression band about 36 ku was found.Conclusion: we obtained OMP2b gene fragment and expressed successfully in Escherichia coli.