Objective To express IL-35 gene in E. coli and purify IL-35 protein. Methods Mouse Ebi3 and p35 mature peptide gene were amplified by RT-PCR from mouse spleen. By overlap PCR, both genes were fused with the ( Gly4Ser)3linker. The fusion gene was inserted initially into pMD19-T vector, then it was subcloned into the expression vector pET-30b ( + ). and was transformed into E. coli BL21 (DE3). The target protein was induced by IPTG and detected by anti-6 x His monoclonal antibody and purified by affinity chromatography. Results Cloned mouse IL-35 gene was identified to contain the expected sequence, the target gene was expressed in the form of fusion protein with relative molecular weight at 50 kd. The protein can be recognized by anti-6 x His monoclonal antibody and can be purified by affinity chromatography. Conclusion Mouse IL-35 was expressed and purified successfully, which lays the foundation for studying the functions and application value of mouse IL-35.
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维普
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