A Novel Vector for Construction of Markerless Multicopy Overexpression Transformants in Pichia pastoris.

ABSTRACT: Pichia pastoris is widely used as a platform for heterologous protein expression because of its high volumetric productivity. Multicopy integration of the target gene is commonly used to improve the production of the target protein. Cre/lox recombination system is a powerful tool for the marker rescue during multiple integrations with one selection marker. Here we reported a novel expression vector based on the Cre/lox recombination system for multiple integrations of target gene to construct multicopy expression strain of P. pastoris. P_AOX1 promoter was fused to cre to construct a methanol inducible Cre recombinase. The leakage expression of Cre recombinase in Escherichia coli was blocked by introducing the operator gene lacO. Th

SUBMITTER: Li D 

PROVIDER: S-EPMC5601908 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature