Description
TEV protease is commonly used site-specific protease that is used for removal of fusion partners and tags from recombinant proteins.
We have constructs for production of both original, wild-type TEV enzyme as well as for activity enhanced variant of it. In both cases the protein is expressed as a fusion with maltose binding protein (MBP) which is cleaved by TEV itself in the expression host, leaving just His-tagged TEV for purification.
Protocol summary
Either of the plasmids are transformed to an E. coli strain for T7 expression system, typically BL21(DE3), for protein production.
Typical preparation of TEV would be expression of 2 liters of culture, followed by purification using Ni-NTA agarose and size-exclusion chromatography.
Materials
pExp-MBP-TEV plasmid for wild type TEV Request
pExp-xMBP-TEVenh plasmid for enhanced activity TEV Request
References