Protein Production Group
About the Group
The COBRE Protein Production Group (PPG) focuses on the cloning, expression and purification of prokaryotic and eukaryotic proteins for COBRE and other investigators in Kansas and the region. The laboratory maintains a variety of equipment to support the production of properly folded proteins in quantities suitable for structural studies (X-ray and NMR), functional studies (catalytic or biological), label-free binding studies (SPR) and/or high throughput (HTP) screening studies.
Laboratory services available include:
- Construct design, cloning, expression and solubility testing
- Protein characterization and analysis
- Large scale expression and purification
- Protein expression in both E. coli and insect cell expression systems
- Protein refolding and reconstitution
- Isotope-labeled proteins (15N, 13C, 2H)
High Throughput Cloning
Ligation Independent Cloning is used to clone genes into a pET Vector without restriction enzyme digestion and ligation. Targeted genes can be cloned in approximately one day with low background. Expressed proteins are purified using one-step affinity chromatography.
Expression of Challenging Proteins
Various fusions are used to increase the expression level, solubility and/or stability of the protein, or to drive the protein into inclusion bodies. The PCR product can be inserted into multiple expression vectors. All tags and fusion proteins are cleavable.