Coloning The Mice Soluble Interleukin 13 Receptorα2 Gene And Expression This Protein In Pichia Yeast Pastoris

In recently years, the incidence of asthma is continuing to rise. During the past decade years, the incidence of asthma increased almost double in Western Europe, and about 15% of adults was suffering from asthma, and about 5% with the patients controled is pool. It is necessary to search a new treatment method about airway inflammation of asthma. Study asthma animal model showed that IL-13 plays an important role in induce airway hyperreactivity (AHR), if have been blocked IL-13 and its signaling pathway ,this is able to promoting the treatment with asthma. The Soluble receptors are important regulators to inflammation and immunity, and its can either down-regulation or up-regulation cytokine signaling and responses, In body fluids, the solubleIL-13Rα2 has been reported to act as an inhibitory protein. it can regulate IL-13 responses, and block IL-13 combined with its membrane receptor. Its can inhibite and reduce treatment of allergenicity to the monoclonal antibody. In currently, sIL-13Rα2 is targeted by the treatment of asthma in mice have no reported. In this study, we will obtain BALB/ c mouse sIL-13Rα2 target gene and recombinant expression vector. the expression vector was transfected into Pichia Yeast pastoris, Obtain recombinant purpose protein.To investigate the application of sIL-13Rα2 treatment asthma and other allergic diseases establish the foundation.Objective: To clone and expression the mice soluble interleukin 13 receptorα2(sIL-13Rα2)gene and Construction the expression vector of the Pichia Yeast pastoris; The expression vector with gene fusion was transfected into GS115 Strains for expression of recombinant sIL-13Rα2 protein. Methods:The totoal 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 cloing target gen by PCR. Target gene was digested with restriction enzyme and cloned into Pichia Yeast expression vector, Recombinant plasmids was transfected into Pichia Yeast pastoris and expressed target protein.and target protein was identificated by SDS-PAGE and Western blot; Results: sIL-13Rα2 target gen was successful cloned by RT-PCR, Target genes have been successfully connecting and constructed recombinant pPICZα-A/sIL-13 Rα2 expression vector. Recombinant expression vector was successful transfected into Pichia Yeast pastoris, sIL-13Rα2 protein was Induced,expression and identification; Conclusions: Obtain BALB/ c mouse sIL-13Rα2 target gene and recombinant expression vector and was transfected into Pichia Yeast pastoris, and Obtained recombinant purpose protein. To investigate the application of sIL-13Rα2 treatment asthma and other allergic diseases establish the foundation.