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Abstract for S30 T7 High-Yield Protein Expression System
The S30 T7 High-Yield Protein Expression System is an E. coli extract-based
cell-free protein synthesis system. It simplifies the transcription and 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 necessary components for translation. This
system can produce high levels of recombinant proteins (up to hundreds of
micrograms of recombinant protein per milliliter of reaction) within an hour.
The investigator supplies only cloned DNA containing a T7 promoter and a
ribosome-binding site (RBS).
The S30 T7 High-Yield Protein Expression System contains the T7 S30 Extract,
Circular, which is prepared from an E. coli strain B deficient in OmpT endoproteinase and lon
protease activity. This results in greater stability for translated proteins that
would otherwise be degraded by proteases if expressed in vivo. An
optimized S30 Premix Plus provides all other required components, including
amino acids, rNTPs, tRNAs, an ATP-regenerating system, IPTG and
appropriate salts to express high levels of recombinant proteins. |
