PCR-based method for identification of integration events in the Pichia pastoris genome

摘要

The yeast Pichia pastoris is a frequently used microorganism for the cloning and expression of eukaryotic genes. These yeast cells have many of the advantages of higher eukaryotic expression systems, such as protein processing, protein folding, and posttranslational modification. P pastoris is a methylotrophic yeast that is capable of metabolizing methanol as its sole carbon source. The promoter regulating the production of alcohol oxidase is used to drive heterologous protein expression in yeast. Stability of recombinant molecules is ensured by homologous recombination between transforming DNA and regions of homology within the genome. Such integrants show extreme stability in the absence of selective pressure.