Yeast Expression Platform
Yeast is a eukaryotic organism and has some advantages and disadvantages for protein expression as compared to E. coli. One of the major advantages is that yeast cultures can be grown to very high densities, which makes them especially useful for the production of isotope labeled proteins. These single-celled eukaryotic organisms grow quickly in defined medium, are easier and less expensive to work with than insect or mammalian cells, and are easily adapted to fermentation. Yeast expression systems are ideally suited for large-scale production of recombinant eukaryotic proteins. crnbio's yeast expression platform provides a flexible, controllable, and effective solution for protein expression needs. Our services includes bioinformatics analysis, domain selection, vector design, tag selection, sequence confirmation, cell transfection, cell amplification, purification by tag selection, yield optimization, scale up processing, and protein characterization.
|
Available Services are as follows, depending on your research needs, you can choose any starting phase or combinations.
Phase I: Subcloning into Expression Vector
|
Description: |
● Amplification/isolation of the gene of interest out of a customer-supplied vector and subcloning it into a suitable expression vector.
● Verification of the subcloned gene by restriction digest and sequencing.
● Preparation of the recombinant vector DNA. |
|
Required Materials: |
● 5 ug of plasmid containing gene of interest plus complete vector sequence in electronic format. |
|
Deliverables: |
● 20 ug of expression-ready recombinant plasmid containing the gene of interest
● Final sequencing data
● Detailed research report |
|
Duration: |
● 2 weeks; If insert > 2kb, one week additional per 1kb |
|
Price (USD): |
● $998; If insert > 2kb, $398 additional per 1kb; |
Phase II: Generation and Identification of Expression Positive Clones
|
Description: |
● Transformation of recombinant constructs into a suitable yeast strain.
● Small scale induction to over-express target protein.
● Test for recombinant protein by SDS-PAGE and western blot (customer must provide appropriate antibodies). |
|
Required Materials: |
● 5 ug of expression-ready recombinant plasmid containing the gene of interest plus complete vector sequence in electronic format. |
|
Deliverables: |
● Expression positive clones
● SDS-PAGE and Western Blot data
● Detailed research report |
|
Duration: |
● 2 weeks |
|
Price (USD): |
● $498 |
|
Optional: |
● For His or GST tagged protein, you can choose to use our antibodies in stock, with and additional charge of $28. |
Phase III: Large-scale Culture
|
Description: |
● One liter of yeast culture will be grown in induction medium suitable for the expression vector used. Recombinant protein expression is harvested for protein purification.
● Test for recombinant protein by SDS-PAGE and western blot (customer must provide appropriate antibodies). |
|
Required Materials: |
● Expression positive yeast clone together with a verified induction procedure |
|
Deliverables: |
● Cell pellets from 1 liter fermentation
● SDS-PAGE or Western Blot data
● Detailed research report |
|
Duration: |
● 1 week |
|
Price (USD): |
● $398 |
|
Optional: |
● For His or GST tagged protein, you can choose to use our antibodies in stock, with and additional charge of $28. |
Phase IV: Protein Extraction & Purification
|
Description: |
● Cell pellet harvested from 1 Liter of yeast culture after induction will be lysized with our optimized reagents and protocol.
● Protein purification using appropriate affinity medium for 6xHis or GST-tagged recombinant proteins.
● Purification can be performed either under native or denatured conditions (as desired by the customer).
● SDS-PAGE and Western Blot will be performed for quality testing. |
|
Required Materials: |
● Cell pellet harvested from 1 liter of yeast culture after induction |
|
Deliverables: |
● Purified protein in standard or customer-specified solution
● SDS-PAGE or Western Blot data
● Detailed research report |
|
Duration: |
● 2-3 weeks |
|
Price (USD): |
● $1500/L, inquire the price for larger volume |
Customer Notices
1. Customer should ensures that the materials provided are correct (including gene templates, primers, expression vectors).
2. Customer is AOR for the mutated gene templates or expression vectors.
References
|
|
| |
