This plasmid allows you to express and purify TEV Protease in your own lab using His-tag affinity purification.
Starting in 1994, scientists began using this protease from tobacco etch virus (TEV) as a means of cleaving and purifying fusion proteins. The sequence for TEV cleavage site can be placed between a protein of interest and an affinity tag. Once the fusion construct has been expressed, it can be bound to beads using the affinity tag. The protein of interest can then cleaved off of the beads, leaving the affinity tag on the beads. Histidine-tagged TEV protease used for cleavage can be removed through incubation with Ni-NTA beads.
This DNA construct contains the 27 kDa catalytic domain of TEV protease fused to the maltose binding protein to increase solubility. The histidine-tagged catalytic domain cleaves itself off of the fusion construct during expression.
TEV protease is regarded as having high and unique specificity, recognizing the cleavage sequence ENLYFQ/X where X is preferably a serine or glycine. It has demonstrated sufficient efficiency at low temperatures (e.g. 4 °C), allowing cleavage while minimizing non-specific proteolytic degradation of target proteins. These advantages have resulted in more frequent use of TEV protease than other commonly available proteases (e.g. factor Xa, thrombin, enterokinase and human rhinovirus 3C protease).
Catalog #: PL0001.TEV