Reconstituted Cell-Free Protein Synthesis System PUREfrex® 2.0
PUREfrex kit is a newly developed reconstituted cell-free protein synthesis reagent based on PURE system technology invented by Professor Takuya Ueda in the University of Tokyo.
The reaction system consists of proteins, ribosome, amino acids and NTPs only (Ref. 1, 2). Those proteins are necessary for transcription, translation and energy regeneration. The proteins and ribosomes are highly purified individually and assembled together to constitute the protein synthesis system. To synthesize your protein, just add your template DNA or mRNA encoding the protein of interest into reaction mixture, and incubate for several hours. This system’s biggest point is RECONSTITUTED system by assembling translation related factors only. By this unique character, you may adjust the composition of reaction mixture as you like and may not have to consider serious background for your downstream application.
By improving the purification process of components of PUREfrex kit, contamination of RNase and β-galactosidase are greatly reduced, in addition to that, lipopolysaccharide (LPS) is also reduced less than 0.1 EU per 1 L of reaction mixture. All proteinous components of PUREfrex kit have no tags for purification and detection. It allows to fuse your protein with any tag to purify the product.
For preparation of following proteins
- prokaryotic protein
- eukaryotic protein
- membrane protein
- protein containing disulfide bonds
- protein containing unnatural amino acids
- ... etc.
For basic research in protein science
- Folding of protein after synthesis
For in vitro display
- Ribosome display
- mRNA display
Feature and Advantages
- No genetic transformation required
- No consideration of cultuire condition required
- No consideration of expression induction condition required
- No consideration of purification condition required
- Cell-free translation reconstituted with purified components. Shimizu et al. (2001) Nat. Biotechnol., vol. 19, p. 751 PMID: 11479568
- Protein synthesis by pure translation systems. Shimizu et al. (2005) Methods, vol. 36, p. 299 PMID: 16076456
- Blanken et al. 2019. Quantitative imaging of gene-expressing liposomes reveals rare favorable phenotypes. Physical Biology. 16(4):1-14. PubMed, Journal
- Bordeau et al. 2014. Curli synthesis and biofilm formation in enteric bacteria are controlled by a dynamic small RNA module made up of a pseudoknot assisted by an RNA chaperone. Nucleic Acids Res. 42(7):4682-96. PubMed, Journal