E. coli S30 Extract System for Circular DNA

The E. coli S30 Extract for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or λ vectors, providing a powerful tool for identifying and characterizing polypeptides. The investigator needs only to supply the cloned DNA containing the appropriate prokaryotic promoter and ribosome binding sites. The S30 Extract for Circular DNA Templates 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 a greater stability of expressed proteins that would otherwise be degraded by pr...

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The E. coli S30 Extract for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or λ vectors, providing a powerful tool for identifying and characterizing polypeptides. The investigator needs only to supply the cloned DNA containing the appropriate prokaryotic promoter and ribosome binding sites. The S30 Extract for Circular DNA Templates 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 a greater stability of expressed proteins that would otherwise be degraded by proteases if expressed in vivo. The S30 in vitro system also allows higher expression levels of proteins that are normally expressed at low levels in vivo due to the action of host-encoded repressors.

Features - Benefits

• Greater Stability: Reduced chance of expressed proteins degrading.
• Complete: Contains all necessary components for coupled transcription/translation.
• Low Background: System 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

• Regulation of prokaryotic gene expression.
• Identification of cloned DNA gene products or mapping of polypeptides to defined DNA fragments.
• Rapid screening for translation from eukaryotic or prokaryotic DNAs cloned in vectors designed for high expression in E. coli.
• Screening of compounds for inhibition of bacterial translation.

For more information, see the Protocols & Applications Guide.

References

1. Zubay, G. (1973) Ann. Rev. Genet. 7, 267–87.
2. Zubay, G. (1980) Meth. Enzymol. 65, 856–77.
3. Pratt, J.M. (1984) In: Transcription and Translation, Hames, B.D. and Higgens S.J., eds., IRL Press, Oxford, 179.
4. Studier, F.W. and Moffatt, B.A. (1986) J. Mol. Biol. 189, 113–30.
5. Collins, J. (1979) Gene 6, 29–42.

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