德国小蠊Bla g 2的原核表达及其多克隆抗体的制备

利用RT-PCR技术扩增德国小蠊Blag2基因,将其克隆进pET-41b载体.在大肠杆菌中经IPTG诱导表达获得GST-Blag2蛋白,SDS-PAGE分析表达产物,得到相对分子量约74kDa的融合蛋白.通过亲和层析法纯化表达的GST-Blag2蛋白,并以此表达产物制成油乳剂疫苗免疫新西兰白兔,制备多克隆抗体.间接ELISA法测得抗体效价达到1∶300000.Western印迹表明抗体有较高的特异性,可分别与GST-Blag2融合蛋白和德国小蠊变应原浸提液反应.Blag2在大肠杆菌中的成功表达及其多克隆抗体的制备,为德国小蠊过敏患者的诊断和治疗提供了帮助.

Prokaryotic Expression of Blattella Bermanica Allergen 2(Bla g 2) and Preparation of Polyclonal Antibody

Bla g 2 gene is amplified by RT-PCR from the total RNA of Blattella germanica and is inserted into pET-41b vector.GST-Bla g 2 was expressed in E.coli via IPTG induction,a relative molecular mass of approximately 74 kDa is shown by SDS-PAGE.The expressed fusion protein is purified by Ni+-agarose affinity chromatography,an emulsion is prepared with the purified Bla g 2 and injected into rabbits.The titer of antiserum identified by indirect ELISA reached 1∶300 000.Moreover,antiserum reacts specifically with purified fusion protein and the extract of Blattella germanica.Preparation of Bla g 2 and its polyclonal antibody offer help to the dialysis and therapy of patients who are allergic to Blattella germanica.