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 deg...

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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.

Features - Benefits

• Flexible: Can translate using any clone that has a T7 promoter and a ribosome binding site. Other S30 extracts require an E. coli promoter.
• Greater Stability: Reduced chance of expressed proteins degrading.
• Complete: Contains all components needed for coupled transcription/translation.
• Low Background: Synthesizes very low levels of endogenous proteins.
• Optimized: Premix is optimized for each lot of S30 Extract and contains all other required components (except amino acids), such as ribonucleotides, tRNAs, PEP (phosphoenol pyruvate) and salts.
• Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/

Applications

• 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.

For more information, see the Protocols & Applications Guide.

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.

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