中文摘要：

为了获得重组Fcε3-4蛋白,从变态反应性哮喘大鼠脾组织中提取总RNA,通过RT-PCR方法克隆大鼠Fcε3-4基因,构建成原核表达载体pET17b-Fcε3-4,并转化大肠杆菌进行诱导表达,Fcε3-4蛋白主要以包涵体形式存在.经Ni-NAT亲和层析柱对尿素溶解的Fcε3-4蛋白进行了纯化,并利用降低尿素梯度的方法柱上对纯化蛋白进行了复性.经Western Blotting和ELISA鉴定,Fcε3-4蛋白具有Fcε的免疫特异性,能被抗大鼠Fcε抗体特异性识别,为下一步研究该蛋白奠定了基础.

英文摘要：

In order to obtain recombinant Fcε3-4 protein the Fcε3-4 gene was amplified by the method of RT-PCR from the total RNA extracted from a fresh spleen of allergic asthma rat, then cloned into expression vector pET-17b. The Fcε3-4 protein was expressed as inclusion body in E. coli BL21. The denatured Fcε3-4 protein was dissolved in 8 mol/L urea,then purified and refolded in lower urea gradient method on Ni-NAT column. The expressed Fcε3-4 owned the epitope recognised by anti-Fcε antibody through Western Blotting and ELISA analysis, which would be a basis for further studies of the Fcε3-4 protein.