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Almost everything you need for PURE-based protein synthesis.

PURESYSTEM® is a novel reconstituted in vitro translation system consisting of about 30 purified enzymes necessary for transcription, translation and energy recycling. All factors for transcription and translation are 6X His-tagged: 3 initiation factors (IF1, IF2, IF3), 3 elongation factors (EF-G, EF-Tu, EF-Ts), 3 release factors (RF1, RF2, RF3), Ribosome recycling factor, 20 aminoacyl-tRNA synthetases, methionyl-tRNA formyltransferase and T7 RNA polymerase. Also included in non-tagged form are E. coli 70S ribosomes, amino acids, NTPs, E. coli tRNA and  an energy recycling system. For template generation we recommend a Protein Express Linear Template PCR kit (use the no His-tag option of the His-tag version or expression plasmids from BioComber (pURE1-4: no tags).

Depicted is the use of PURESYSTEM® to synthesize and purify untagged target proteins from linear PCR-based templates or expression plasmids. We recommend expression plasmids from BioComber (pURE1-4).

Example of expression and reverse purification of DHFR produced in PURESYSTEM®

Examples of fluorescent lysine-labeled proteins produced in PURESYSTEM®. Proteins were separated on a 10-20% gradient SDS PAGE gel, stained with SYPRO® Red and analyzed on a fluorescent gel scanner. Red bands: PURESYSTEM® component proteins. Green bands: Synthesized target proteins.

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