PEPCF expresses proteins in bacteria, insect and mammalian cells and uses a variety of chromatographic and biophysical techniques for protein purification and characterization.
Overview of our insect cell expression vectors
Please note that only the vectors indicated in yellow can be shared with external users via the EMBL MTA.
We also have a collection of insect cell expression vectors specifically adapted for ligation-independent cloning. These vectors are called pCoofy vectors and were developed by Sabine Suppmann at the Max Planck Institute for Biochemistry in Martinsried, Munich.
The detailed cloning protocols and descriptions of these vectors can be found in the following reference:
Scholz J., Besir H., Strasser S. and Suppmann S. (2013) A new method to customize protein expression vectors for fast, efficient and background free parallel cloning. BMC Biotechnology. 13:12
Internal EMBL users can obtain these vectors at the Protein Expression and Purification Core Facility. External users can request them via Addgene.
| Name | N-terminal tag | C-terminal tag | Antibiotic resistance |
|---|---|---|---|
| pCoofy27 | His7-3C | X | Ampicillin Gentamycin |
| pCoofy28 | His6-GST-3C | X | Ampicillin Gentamycin |
| pCoofy29 | His6-MBP-3C | X | Ampicillin Gentamycin |
| pCoofy41 | 3C (not expressed) | * His10 * TwinStrepII * S-tag * CBP | Ampicillin Gentamycin |
| pCoofy51 | TwinStrepII-3C | X | Ampicillin Gentamycin |
| pCoofy59 | TrxA-His6-3C | X | Ampicillin Gentamycin |
| pCoofy60 | His6-eGFP-3C | X | Ampicillin Gentamycin |
| pCoofy63 | His6-SumoStar (Smt3 R64T R71E) | * none * His10 * TwinStrepII * EPEA-tag | Ampicillin Gentamycin |
| pCoofy64 | GP67ss-His6-SumoStar (Smt3 R64T R71E) | * none * His10 * TwinStrepII * EPEA-tag | Ampicillin Gentamycin |
For the expression of protein complexes in insect cells, we have both the MultiBac and biGBac vectors available.
More information about how to use the MultiBac system can be found on the websites of Geneva Biotech and the University of Bristol.
A detailed description of the biGBac system can be found in the following references:
Weissmann F., Petzold G., VanderLinden R., Huis in ‘t Veld P.J., Brown N.G., Lampert F., Westermann S., Stark H., Schulman B.A. and Peters J.-M. (2016) biGBac enables rapid gene assembly for the expression of large multisubunit protein complexes. PNAS. 113(19): E2564-2569
Weissmann F. and Peters J.-M. (2018) Expressing Multi-subunit Complexes Using biGBac. Methods Mol Biol. 1764: 329-343