Multicopy plasmid integration in Komagataella phaffii mediated by a defective auxotrophic marker.

ABSTRACT:

Background

A commonly used approach to improve recombinant protein production is to increase the levels of expression by providing extra-copies of a heterologous gene. In Komagataella phaffii (Pichia pastoris) this is usually accomplished by transforming cells with an expression vector carrying a drug-resistance marker following a screening for multicopy clones on plates with increasingly higher concentrations of an antibiotic. Alternatively, defective auxotrophic markers can be used for the same purpose. These markers are generally transcriptionally impaired genes lacking most of the promoter region. Among the defective markers commonly used in Saccharomyces cerevisiae is leu2-d, an allele of LEU2 which is involved in leucine metabolism. Cells transformed with this marker can reco

SUBMITTER: Betancur MO 

PROVIDER: S-EPMC5465527 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature