Restriction digests of the clone give the following sizes (kb): HindIII--5.0; BamHI--5.0; EcoRI--5.0; XbaI--5.0. Expression vector permitting production of a fusion protein. The glutathione S-transferase (GST) fusion protein can be purified by glutathione affinity chromatography, and the desired polypeptide released from the fusion product by thrombin. Constructed from pGEX-2T by inserting an oligonucleotide at the EcoRI site which encodes the glycine "kinker" and additional restriction sites to facilitate cloning in all reading frames. The order of the major features in this plasmid are: Ptac - GST - thrombin cleavage site - BamHI site - SmaI site - glycine "kinker" - multiple cloning site - ampR - pBR322 ori - lacIq. |
