E. coli T7 S30 Extract System for Circular DNA

The E. coli T7 S30 Extract System for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or λ vectors containing a T7 promoter by providing an extract that contains T7 RNA polymerase for transcription and all components needed for translation. The investigator only supplies cloned DNA containing a T7 promoter and a ribosome binding site. This product is prepared by modifications of the method described by Zubay from an E. coli strain B deficient in OmpT endoproteinase and lon protease activity. This results in greater stability of expressed proteins that would otherwise be degraded by proteases if expressed in vivo.

The E. coli T7 S30 Extract System for Circular DNA enables you to translate using any clone that has a T7 promoter and a ribosome binding site; No E. coli promoter required. Proteins expressed with this system demonstrate reduced tendency to degrade and have shown very low levels of endogenous protein.

For more information, see the Protocols & Applications Guide.

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• Synthesis of radiolabeled protein.
• Verification of cloned gene expression before in vivo protein expression in E. coli.
• Use of protein in functional studies of transcription and translation.
• Use of protein as a tracer in protein purification.
• Incorporation of unnatural amino acids into proteins for structural studies.

References

1. Nevin, D.E. and Pratt, J.M. (1991) FEBS Lett. 291, 259–63.
2. Zubay, G. (1973) Ann. Rev. Genet. 7, 267–87.
3. Zubay, G. (1980) Meth. Enzymol. 65, 856–77.
4. Studier, F.W. and Moffatt, B.A. (1986) J. Mol. Biol. 189, 113–30.
5. Pratt, J.M. (1984) In: Transcription and Translation, Hames, B.D. and Higgens S.J., eds., IRL Press, Oxford, 179.