中文摘要：

目的原核表达并纯化小鼠白细胞介素-13受体α2胞外区（sIL-13Rα2）。方法将sIL-13Rα2基因克隆入原核表达载体pROEXHTa,构建重组表达质粒pROEXHTa/sIL-13Rα2,经双酶切鉴定后,转化大肠杆菌BL21（DE3）,IPTG诱导表达。薄层扫描分析目的蛋白的表达量,并对其进行表达形式分析及Westernblot鉴定。镍柱亲和层析纯化目的蛋白。结果重组表达质粒经双酶切鉴定证明构建正确。表达的sIL-13Rα2蛋白相对分子质量约为36200;诱导6h,重组蛋白的表达量最高,约占菌体总蛋白的45%;重组蛋白以包涵体形式存在,可与大鼠抗小鼠sIL-13Rυ2单克隆抗体发生特异性反应;纯化后纯度大于95%。结论已成功表达并纯化了sIL-13Rα2,为进一步探讨其治疗支气管哮喘奠定了基础。

英文摘要：

Objective To express mouse IL-13 receptor α2 extracellular domain（sIL-13Rα2）in prokaryotic cells and purify the expressed product. Methods The sIL-13Rα2 gene was cloned into prokaryotic expression vector pROEX HTa, and the constructed recombinant plasmid pROEX HTa / sIL-13Rα2 was identified by restriction analysis, then transformed to E. coli BL21 （DE3）for expression under induction of IPTG. The expressed product was determined for expression level by thin layer scanning, analyzed for form, and identified by Western blot, then purified by nickel ion affinity chromatography. Results Restriction analysis proved that recombinant plasmid pROEX HTa / sIL-13Rα2 was constructed correctly. The expressed sIL-13Rα2 protein, with a relative molecular mass of about 36 200, reached a maximum expression level after induction for 6 h, which contained about 45% of total somatic protein. The recombinant sIL-13Rα2 protein, existing in a form of inclusion body, showed specific reaction with monoclonal antibody against mouse sIL-13Rα2, and reached a purity of more than 95% after purification. Conclusion Mouse sIL-13Rα2 was successfully expressed and purified, which laid a foundation of further study on its application to treatment of bronchial asthma.