pSparkleTEV is an open source plasmid allowing the user to express the protease from tobacco etch virus (TEV) in E. coli. TEV protease is commonly used to remove purification tags from other recombinant proteins. It is highly sequence-specific, recognizing the consensus sequence "ENLYFQ/G" present in many pSparkle plasmids.
Like all plasmids in the pSparkle series, pSparkleTEV is optimized for expression in our cost-effective and cool-looking SparkleCell desktop bioreactors. It may work in other systems, such as shaker flasks, although such use will be less efficient and will likely require optimization of conditions.
TEV protease is expressed as an MBP fusion protein with his-tag. These tags are not removable. They are helpful both for purifying your TEV protease for the first time, and for removing it from a cleavage reaction later.
In our hands, the yield of MBP-TEV protease protein (after purification) is typically 300 mg per run in SparkleCell EasyGrow and 2.3 grams in SparkleCell PowerGrow. Under realistic conditions (substrate fusion protein with two bulky groups on both sides of the cleavage site, provided at a concentration of 1 mg/ml), each MBP-TEV molecule will cleave approximately one substrate per hour:
We offer ready-made, purified MBP-TEV protease protein as well. This is the same protein encoded by pSparkleTEV. The TEV protein product's web page page has more information on how the protease is used.