Expression of Human GLP-1-IgG Fc Fusion Protein in Pichia pastoris

Objective:Construct and high-effectively express the fusion protein GLP-1-IgG Fc in the Pichia pastoris.Method:Modified the sequence of GLP-1 by eliminating its peptide enzymes sites.The chimeric gene GLP-1-IgG Fc achieved through gene splicing by overlap extension(SOE) method was cloned into pPIC9K and transformed into Pichia pastoris.The fusion protein was expressed in Pichia pastoris and analyzed using SDS-PAGE and Western Blot.Result:The chimeric gene GLP-1-IgG Fc and yeast expression vector pPIC9k/GLP-1-IgG Fc were constructed.The fusion protein GLP-1-IgG Fc was successfully expressed in the Pichia pastoris and the expression level of fusion protein in 1 L flask reached 5 mg/L after 72h growth.The results of SDS-PAGE and Western-Blot showed the expressed protein was the GLP-1-IgG Fc fusion protein.Conclusion:The human GLP-1-IgG Fc fusion protein was expressed successfully in Pichia pastoris,and it could be the foundation for the GLP-1-IgG Fc activity and half-life determination.