Expression and purification of TEV protease

The plasmid (pSF455) encodes a modified TEV as a fusion protein with MBP in pQE80N (MBP-tev-H14-TEV(CDS)-Arg5). The original TEV contains a self-cleavage site that has been removed by S Frey. The Arg tail served for better purification by an ion exchange step. The MBT is cleaved off in vivo.

Buffer:

extraction buffer:

50 mM K-PO4 [pH 7.5], 300 mM NaCl, 1 mM Imidazol

elution buffer:

extraction buffer, 300 mM imidazol

storage buffer:

20 mM Na-phosphate [pH 7], 50 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 10% glycerol