中文摘要：

目的：构建GM-CSF原核表达载体,并诱导表达、纯化其蛋白。方法：将GM-CSF基因克隆至原核表达载体pET-32中,转化至BL21中,用智能蛋白质多维纯化系统（AKTAxpressTM）纯化目的蛋白,利用MTT法测定GM-CSF融合蛋白对绵羊外周血淋巴细胞的增殖活性。结果：成功获得了435bp的绵羊GM-CSF基因片段,诱导表达了35kDa融合蛋白GM-CSF,其纯度达95%以上,浓度达860mg/mL;通过MTT证明该融合蛋白对绵羊淋巴细胞具有增殖活性,其作用最明显的蛋白浓度是200μg/mL。结论：成功获得重组蛋白GM-CSF,该蛋白具有较好的生物活性,为免疫增强佐剂的开发奠定了基础。

英文摘要：

Objective： Cloning, expression and purification of sheep recombinant GM -CSF. Method：The gene of GM -CSF was cloned in- to pET32 vector,and expressed in Escherichia coli strain BI21. The recombinant protein was purificated by AKTAxpressTM ,MTT method was adopted to proliferative activity of GM - CSF which could stimulate proliferation of sheep peripheral blood iymphocytes. Result： Its suc- cossful to clone a GM - CSF cDNA of 435bp into pET32a vector, and express recombinant GM - CSF of 35kDa induced in E. coli. Its purity highed than 95% and concentration of 860mg/mL. it has been identified by MTr that Fusion protein had an proliferative activity on culti- vating lymphocytes of sheep, and the most efficacious concentration is 200μg/mL. Conclusion：The consequences manifest that purity G M -CSF with good bioactivity was gained and provided an important basis for immune -enhancing adjuvant.