Introduction to multi-copy Pichia expression vectors
The Invitrogen Multi-Copy Pichia Expression Kit (Cat. no. K1750-01) is designed for the isolation and expression of recombinant proteins from Pichia pastoris strains that contain multiple copies of a specific gene. The kit facilitates protein production in this methylotrophic yeast, which offers advantages such as eukaryotic post-translational modifications and high heterologous protein expression levels compared to E. coli or Saccharomyces cerevisiae.
The manual details the experimental outline for using the kit, including methods for cloning into multi-copy vectors (pPIC3.5K, pPIC9K, pAO815), transformation of Pichia strains via spheroplast preparation and electroporation, in vivo screening of multiple inserts, and optimization of protein expression. It also covers downstream processing steps such as SDS-PAGE analysis, scale-up procedures, and protein purification.
| Kit Contents: Vectors Storage Temperature | -20°C |
|---|---|
| Kit Contents: Media Yield (YP Base Medium) | 2 liters per pouch |
The kit is designed for the isolation and expression of recombinant proteins from Pichia pastoris strains.
The Spheroplast Module contains enough reagents for 10 preparations.