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The yeast protein expression system is a cost-effective eukaryotic expression platform capable of both secretion and intracellular expression. Exogenous genes expressed in yeast undergo post-translational processing, resulting in folded proteins with glycosylation modifications.
These proteins exhibit greater stability compared to those expressed in prokaryotic systems, making the yeast system particularly suitable for expressing eukaryotic genes and producing functional proteins. In yeast secretion expression, proteins are secreted into the extracellular matrix, facilitating the production of high-purity proteins.
Service Process
1. Expression vector construction
Process
- Codon optimization, gene synthesis
- The PCR amplification products are ligated to the expression vectors e.g. pPic9k, pPic3.5k, pPiczαA, etc.
- Transform ligation mixtures into E. coli strain
- Obtain the correct recombinant plasmid
Features
Multiple vectors optimisation - To enhance expression success and achieve higher yields, apart from conventional N-terminal fusion protein expression, we also offer proteins with C-terminal fusion labels
Lead Time: 15-20 business days
2. Transformation and strain identification
Process
- Prepare the recombinant plasmid in large quantities
- Linearization of recombinant plasmid
- Transform to GS115, X33, KM71 and other hosts by electroporation
- PCR analysis is recommended to verify successful transformants
- Use geneticin G418, Zeocion and other antibiotics for multiple copies screening to obtain high copy
Lead Time: 10-15 business days
3. Small test, scale up expression and purification
Process
- Small scale expression screening (20-40 strains)
- Determine strain and optimise expression conditions
- Scale up culture and protein purification
Features
Multiple purification methods (optional) - Determine the optimal purification method, using different chromatographic conditions
Lead Time: 10-15 business days
4. Optional additional services
Process
- Endotoxin removal, lyophilisation or filter-sterilisation
- Tag removal (Additional charge)
Features
Customers have the flexibility to select from a range of additional services tailored to their specific requirements
Lead Time: 5 business days
6. Quality control
Process
- The purity, concentration and further tests are performed.
Features
Detailed COA report is provided.