Effect of Plasmid Design and Type of Integration Event on Recombinant Protein Expression in Pichia pastoris.

ABSTRACT: Pichia pastoris (syn. Komagataella phaffii) is one of the most common eukaryotic expression systems for heterologous protein production. Expression cassettes are typically integrated in the genome to obtain stable expression strains. In contrast to Saccharomyces cerevisiae, where short overhangs are sufficient to target highly specific integration, long overhangs are more efficient in P. pastoris and ectopic integration of foreign DNA can occur. Here, we aimed to elucidate the influence of ectopic integration by high-throughput screening of >700 transformants and whole-genome sequencing of 27 transformants. Different vector designs and linearization approaches were used to mimic the most common integration events targeted in P. pastoris Fluorescence of an

SUBMITTER: Vogl T 

PROVIDER: S-EPMC5835746 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature