pGEX2T-27k is a pBR322 derived plasmid that is AmpR and has medium copy number. Expression of TEV protease is induced by IPTG. We recommend replacing tryptone in rich medium by NZ amine A because the latter does not contain lactose which induces the tac promoter. The GST-TEV protease hybrid is expressed as an inclusion body in the presence of high IPTG. Soluble (and active) TEV protease is best obtained at 10 µM IPTG.

- Reference:
Parks, T., K. Leuther, E. Howard, S. Johnston, and W. Dougherty. 1994. Release of proteins and peptides from fusion proteins using a recombinant plant virus proteinase. Anal. Biochem. 216: 413-417.

183-211 Ptac
258-930 GST
930-1671 TEV protease fragment
2070-2930 bla
4011-5093 lacI^Q

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