TEV protease expression vectors

TEV NIa protease as a tool for in vitro and in vivo proteolysis

	Databases ECCE
	Maltose transporter
	DNA sequences
	Software Dana's tools
	Strategy Gene expression in E. coli
	TEV NIa protease as a tool

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2. TEV protease expression vectors

For in vivo proteolysis
Medium or low copy number vectors such as pBR322, pACYC184 or pHSG575 are recommended. To limit TEV protease levels under non-inducing conditions tight promoters are required. Lac or Tet promoters can be used if lac or tet repressor genes are on the plasmid.
TEV protease does not lose its activity if it is part of a hybrid protein.
For purification of TEV protease
Follow the suggestions for gene expression in E. coli. The choice of vector system is not so critical. T7 polymerase controlled expression vectors are often used. Growth at 20oC will lead to mainly soluble TEV protease while growth at 37oC will produce inclusion bodies. Inclusion bodies are not a problem because TEV protease can be efficiently refolded. N-terminal His tags facilitate purification and do not interfere with activity or refolding.