The expression of murine soluble interleukin 13 receptor α2 and initial activity analysis in Pichia Yeast pastoris

Objective:To clone and express murine soluble interleukin 13 receptor α2(sIL-13Rα2)gene in Pichia Yeast pastoris.Methods:The total RNA extracted from spleen cells of BALB/c mouse as template,sIL-13Rα2 cDNA enconding extracellular domain of sIL-13Rα2 chain was obtained by RT-PCR.Using cDNA template the cloned target gene was amplified by PCR.Target gene was digested with restriction enzyme and cloned into Pichia Yeast expressing vector.Recombinant plasmids was transfected into Pichia Yeast pastoris for expressing target protein,and the target protein was identificated by SDS-PAGE and Western blot.Results:sIL-13Rα2 target gene was successfully cloned into the recombinant pPICZα-A/sIL-13Rα2 expression vector.The recombinant expressing vector was successfully transfected into Pichia Yeast pastoris,and sIL-13Rα2 protein was positively expressed by identification.Conclusion:Murine sIL-13Rα2 target gene is obtained and recombinantly expressed into the Pichia Yeast pastoris vector.It may establish the foundation for further application of sIL-13Rα2 in treatment asthma and other allergic diseases.