Description:
pTYB2 is an E. coli cloning and expression vector (7474 bp) used in the IMPACT™ Kit (NEB #E6901) which allows the overexpression of a target protein as a fusion to a self-cleavable affinity tag (1,2). This C-terminal fusion vector is designed for the in-frame insertion of a target gene into the polylinker upstream of an intein tag (theSce VMA intein/chitin binding domain, 55 kDa) (1,2). This results in the fusion of the C-terminus of the target protein to the N-terminus of the intein tag. Thiol-induced self-cleavage of the intein releases the target protein from the chitin-bound intein tag, resulting in a single column purification of the target protein.
For fusion of the N-terminus of the target protein to the intein tag, use pTYB11 (NEB #N6901) or pTYB12 (NEB #N6902). This vector can be used in conjuction with pTYB12 (NEB #N6902) to test which fusion construction (N-terminal or C-terminal) maximizes the expression and yield of a target protein (3).
Source:
pTYB2 is isolated from an E. coli strain (r-m-) by a standard plasmid purification procedure.
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