The protein expression unit at the Structure Proteomics Unit expresses and produces recombinant proteins from various sources.
Target genes of interest cloned in appropriate expression vectors are introduced into host cells to produce recombinant proteins. To increase the probability that a given protein will be expressed in a soluble and active form, and in high yield, several expression systems are used. These include bacterial systems and three eukaryotic systems - yeast, insect and mammalian cells.
Small scale expression tests:
- Small scale expression test screening (4-50 ml cultures) in E. coli using 2 induction temperatures or 2 bacterial strains or 2 different fusion tags.
- Small scale expression test screening (10-50 ml cultures) in P. pastoris.
- Small scale expression test screening (4-30 ml cultures) in Sf9 and Tni.
- Small scale expression test screening (4-20 ml cultures) in HEK293.
- Large scale production (1.5-5L) in the appropriate expression system in optimized conditions.
- Analysis of soluble and insoluble cellular fractions by SDS-PAGE for protein expression and Western Blot with appropriate antibodies, if necessary.