Yeast Expression System
Yeast is a eukaryotic organism and has some advantages and disadvantages compared with E. coli. One of the main advantages is that yeast cultures can grow to very high densities and are suitable for production of isotope-labeled proteins for NMR. The two most common used yeast strains are Saccharomyces cerevisiae and the methylotrophic yeast Pichia pastoris. Various yeasts are extremely useful for the expression and analysis of eukaryotic proteins. These yeast strains have been well characterized genetically and are known to undergo many post-translational modifications. These single-celled eukaryotes grow rapidly in a specific medium, which are easier to use and lower in cost than insect or mammalian cells.
Overall solution
- Yeast Scale-up Expression
- Vector construction, plasmid production, and strain creation
- 50 ml small scale expression analysis with affinity pulldown
- 1 liter expression, and one-step affinity purification
- 1 liter expression, and tandem affinity purification
Workflow
1. Clone the target gene into the pJ expression plasmid (Biogrammatics) with N- or C-terminal His and/or Strep tags and the n-terminal α mating factor secretion signal peptide, which can produce up to 100 ug plasmid DNA.
2. Introduce the DNA into the yeast strain, select clones so that the target DNA is stably integrated on the antibiotic-containing medium.
3. Perform small-scale expression analysis of the target protein in two or more clones, collect samples of cells and conditioned medium every day. The conditioned medium samples were analyzed by PAGE and Western blotting, and the clone and time point showing the highest expression of secreted protein were selected for affinity pulldown analysis. Affinity pulldown was performed from the conditioned medium and analyzed by PAGE and Western blotting.
4. Under the expression conditions determined in the vial expression analysis, perform 1 liter expression in a shake flask and harvest. The protein can be purified by affinity chromatography using nickel resin or tandem affinity chromatography using nickel and chain actin resin, with or without SEC. The eluted fractions are analyzed by PAGE and Western blotting, combined according to the purity indicators established with the customer.
Our advantages
- CD BioSciences flexibly and comprehensively provide all services related to Yeast expression system, including small-scale expression and purification, solution optimization and large-scale expression.
- Our research supervisor and project manager will complete your project on time through open communication channels and meet your requirements.
- CD BioSciences has established a professional after-sales service team to provide customers with efficient and practical solutions.
Our service items
| Project name | Yeast expression system |
|---|---|
| Our services |
|
| Sample requirements | DNA encoding target protein |
| Product delivery mode | We will complete the project on time according to the agreed time. CD BioSciences offer you the following results: 1. Purified protein 2. Study report 3. Certificate of Analysis |
| Price | Inquiry |
Why choose us?
CD BioSciences provides professional yeast expression systems to meet the needs of regular customers to determine hits on time and on budget. CD BioSciences relies on the world-class protein expression platform and technical expertise, we provide customers with the best quality one-stop Yeast expression systems, including the development of experimental procedures according to different experimental needs. Please feel free to contact us for more detailed information, our scientists will tailor the most reasonable plan for your project.
