中文摘要：

目的克隆并在毕赤酵母中表达小鼠sIL-13Rα2基因。方法提取小鼠脾脏细胞总RNA，逆转录为cDNA，应用分段PCR的方法将sIL-13Rα2基因定向克隆至酵母表达质粒pPICZα—A，并转染至毕赤酵母GS115（his4）菌株，进行目的蛋白的诱导表达。表达的蛋白经SDS—PAGE和Western blot，进行分析。结果重组表达质粒pPICZα—A／sIL-13Rα2经酶切鉴定，表明目的基因为正向插入。重组酵母菌的PCR鉴定结果显示，目的基因片段已插入酵母染色体中。目的蛋白表达量约占菌体总蛋白的30％，每升发酵液约含2mg重组蛋白。重组蛋白可与兔抗小鼠sIL-13Rα2单抗发生特异性反应。结论已成功克隆并在毕赤酵母中表达了小鼠sIL-13Rα2基因，为应用sIL-13Rα2蛋白治疗哮喘及其他过敏性疾病奠定了基础。

英文摘要：

Objective To clone murine soluble interleukin-13 receptor α2 （sIL-13Rα2） gene and express in Pichiapastoris. Methods Total RNA was extracted from routine spleen cells for amplification of sIL-13Rα2 gene by RT-PCR. The amplified gene fragment was cloned into expression vector pPICZα-A, and the constructed recombinant plasmid pPICZα-A/sIL-13Rα2 was transfected to P. pastoris GS115 （his4） for expression under induction of methanol. The expressed protein was identified by SDS-PAGE and Western blot. Results Restriction analysis proved that the target gene was inserted forwardly into recombinant plasmid pPICZα-A/ sIL-13Rα2. PCR proved that the target gene was inserted into the chromosome of recombinant P. pastoris. Each liter of fermentation broth contained 2 mg of expressed protein. The expressed product contained about 30% of total somatic protein and showed specific reaction with rabbit anti-mouse slL-13Rα2 McAb. Conclusion Murine sIL-13Rα2 gene was successfully cloned and expressed in P. pastoris, which laid a foundation of treatment of asthma and other allergic diseases with slL-13Ra2 protein.