muc_1100的表达、纯化及活性探究

Objective: In this study, we aimed to construct a prokaryotic expression plasmid of gene Amuc_1100 and employ ELP(elastin-like protein) as a purification tag for fusion expression, and explore its biological activity. Methods: Design and synthesize the recombinant plasmid pET28a-Amuc_1100-ELP and transform into E.coli BL21(DE3), the recombinant protein was purified by the inverse transition cycle characteristic of ELP. In order to detect the biological activity of the recombinant protein, the expression level of interleukin-10 (IL-10) was measured in PBMCs cells from mice, and the effect of the protein on the metabolism of mice was observed by oral administration in HFD-induced mouse model. Results: These results showed that molecular weight of the expressed protein was 60 kDa as expected. The high purity recombinant protein was obtained after inverse transition cycling (ITC). Moreover, the recombinant protein promoted the expression level of IL-10 and improved the HFD-induced metabolic disorders. Conclusion: The prokaryotic recombinant plasmid pET28a-Amuc_1100-ELP was successfully cloned, a high purity and active target protein was obtained after expression and purification. Taken together, our findings provide support for the industrial production and clinical application.