牛布鲁氏菌OMP25的原核表达及表达产物的免疫学特性

Prokaryotic Expression of B.abortus OMP25 and ImmunologicalCharacteristics of the Expressed Products

目的研究布鲁氏菌外膜蛋白OMP25的克隆、测序和表达,并对其进行免疫特性检测。方法采用PCR技术对牛布鲁氏菌外膜蛋白OMP25基因进行了扩增和克隆测序,并成功构建了原核表达质粒pGEX-OMP25。将其转入E.coliBL21,经IPTG诱导表达,用电洗脱纯化后获得了高纯度的目的蛋白。纯化后的蛋白与弗氏佐剂乳化后免疫家兔得到了较高效价的抗血清。结论通过ELISA、Western-blotting分析及特异性检测试验,证明目的蛋白不仅具有抗原性且有良好的免疫反性。

Objective To clone and express the bmcella outer membrane protein OMP25 and then detect its reactivity to the Bmcella immunized rabbit serum. Mehtods The B.abortus A544 gene was PCR amplified and sequenced, and the prokaryotic expression plasmid pGEX-OMP25 successfully constructed. The plasmid was transferred to the E.coli BL21, and induced with IPTG. High purity protein was obtained by power elution purification. When the purified protein emulsified with Freund＇s adjuvant was used to immunize rabbits, higher antiserum titer was obtained. Conclusion It was shown by ELISA, Western-blotting analysis and specificity testing, that the protein had not only a good antigenicity, but had also a high immunogenicity.