Expression of Human Serum Albumin and Interferon α2b Fusion Protein in the PichiaPink System
Objective: The aim of this experiment was to compare the differences between the PichiaPink strain and Pichia pastoris GS115 strain, and further explore the potential advantages of the PichiaPink system. Methods: The recombinant expression vectors contained the fusion gene coding human serum albumin and interferon α2b fusion protein (HAS-IFN-α2b) were constructed, and were transformed them into the P.pastoris GS115 and the PichiaPink strain respectively. The fusion protein of HAS-IFN-α2b were expressed by methanol induction, and the expression level was evaluated. The SDS-PAGE analysis was used to detect the degradation of HAS-IFN-α2b in the P.pastoris GS115 and the PichiaPink strain. Results: Essentially all transformants in the PichiaPink system showed to express the objective protein of HAS-IFN-a2b, but only 60% transformants in the GS115 strain displayed to express the target protein. In the same kind strains of Pink, the expression levels of HAS-IFN-α2b were respectively compared, which discovered the strain that have integrated with the pPink-HC vector is higher expression than the strain that have integrated with the pPink-LC vector. The PichiaPink strains which had knockouted three proteases genes expressed the fusion protein of HAS-IFN-α2b to exhibit a little degradation in YPD and BMMY medium, but HAS-IFN-α2b degradation phenomenon showed very serious in BSM medium. Conclusion: The results have indicated to have an important value for high-level expression and large-scale production of secreted recombinant proteins in PichiaPink system.
PichiaPink systemPichia pastorishuman serum albumin and interferon α2b fusion protein
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NSTL
万方数据
维普
