Engineering the pPICZαA Vector for Optimized Recombinant Protein Expression in Pichia pastoris: Substitution of Antibiotic Resistance and Auxotrophic Selection Markers

Abstract

Background: Plasmids play a crucial role in biotechnology by facilitating the transfer and manipulation of heterologous DNA in host cells for the production of recombinant proteins. The pPICZαA plasmid, featuring the AOX1 promoter, is regarded as one of the best expression vectors in Pichia pastoris strains. However, the product of the zeocin antibiotic resistance gene (BleR) can be toxic to the host during stable growth, and the high cost of this antibiotic presents a significant barrier to the use of this plasmid.