PUREfrex 1.0

Cosmo Bio LTD

Catalog No: GFK-PF001-0.25-EX

Explore all PUREfrex® products and accessories from our PUREfrex® homepage

PUREfrex Products: Quick Access

Kits	Supplements
PUREfrex 2.0 PUREfrex 2.1 PUREfrex 1.0	DnaK Mix GroE Mix DsbC Set PDI Set EF-P

PUREfrex® (manufactured by Gene Frontier Corporation) is an E. coli reconstituted coupled transcription/translation system based on the PURE system originally developed by the Ueda group (Shimizu et al., 2001) and later commercialized as PURESYSTEM® (Shimizu and Ueda, 2010) by BioComber (Tokyo Japan). PUREfrex® contains highly purified E. coli proteins comprising modules for aminoacylation, energy regeneration, transcription and translation. Among the many advantages of PUREfrex® over competing coupled transcription/translation systems are reduced levels of contaminating proteases, nucleases, and phosphatases, beta-galactosidase and LPS. Compared to cell lysate-based systems, the more defined and modular chemistry of PUREfrex® leads to greater yield, reproducibility and flexibility. The untagged protein components of PUREfrex® (in contrast to the hexahistidine-tagged components of BioComber's PURESYSTEM® and New England Biolab's PURExpress® system) permit addition of any desired tag (including His-tags) to a protein of interest and subsequent rapid and simple purification by a single immobilized metal affinity chromatography step. Protein yields can reach 600ng/ul.

PUREfrex® is available in three forms: PUREfrex® 1.0, PUREfrex® 2.0 and PUREfrex® 2.1.

PUREfrex® can be used to express proteins containing non-natural amino acids by exploiting Protein Express's  CloverDirect™ line of tRNA reagents for site-directed protein functionalization. CloverDirect™ UAG or CGGG aminoacyl tRNAs are available charged with different non-natural amino acids that confer a range of functionality, including fluorescent-labeling, biotin-labeling, PEGylation, chemical crosslinking and photo-regulation. Targeting incorporation of specific non-natural amino acids is achieved by designing protein expression templates with UAG and/or CGGG codons at user-defined positions.

For template construction, we recommend Protein Express Linear Template PCR kits (available in two forms: His-tag only or ProX-tag + His-tag) or expression plasmids from Protein Express (pROX-FL92.1: ProX-tag + His-tag) or BioComber (pURE1-4: no tags). The ProX tag is a special N-terminal peptide designed for the incorporation of certain large unnatural amino acids that require precise N-terminal positioning (codon 9) and a specific codon context (flanked by AAC and TCT). Codon 9 of the ProX tag can be varied between UAG, CGGG or TTT (as a control).

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