The E. coli T7 S30 Extract System for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or lambda vectors containing a T7 promoter.
- Create Radiolabeled Protein for Transcription/Translation Studies
- Translate using clones with a T7 promoter and a ribosome binding site; no E. coli promoter required
- Reduce the chance of expressed proteins degrading
- Includes all components needed for coupled transcription/translation
The E. coli T7 S30 Extract System for Circular DNA simplifies the transcription/translation of DNA sequences cloned in plasmid or lambda 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.
Applications include 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 and as a tracer in protein purification, and incorporation of unnatural amino acids into proteins for structural studies.
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.
