This is a product manual that introduces pGAPZ A, B, C and pGAPZα A, B, C Pichia expression vectors for constitutive expression and purification of recombinant proteins.
This manual provides instructions for using the pGAPZ A, B, and C and pGAPZα A, B, and C Pichia expression vectors (Catalog nos. V200–20 and V205–20) for the constitutive expression and purification of recombinant proteins in Pichia pastoris. The vectors utilize the GAP promoter to express proteins fused to a C-terminal myc epitope and polyhistidine tag, or an N-terminal α-factor secretion signal, depending on the specific vector used.
The document details the experimental process, including cloning strategies, transformation into E. coli, and transformation into Pichia pastoris strains such as X-33, GS115, KM71, and SMD1168. It covers methods for optimizing protein expression, purification using metal-chelating resin (e.g., ProBond™), and technical considerations for recombination and integration.
| Vector Size (pGAPZ A, B, and C) | 2.9 kb |
|---|---|
| Vector Size (pGAPZα A, B, and C) | 3.1 kb |
| Promoter Used | GAP promoter |
| Selectable Marker | Zeocin™ |
| Storage Temperature (Vectors) | -20°C |
| Storage Temperature (Stabs) | 4°C |
Vectors should be stored at -20°C immediately upon receipt.
They produce proteins fused to an N-terminal peptide encoding the Saccharomyces cerevisiae α-factor secretion signal.