Cloning of Murine Soluble Interleukin-13 Receptor α2 Gene and Its Expression in Pichia pastoris

Objective To clone murine soluble interleukin-13 receptor α2(sIL-13Rα2)gene and express in Pichia pastoris.Methods Total RNA was extracted from murine 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 transfect-ed 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 sIL-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 sIL-13Rα2 protein.