What Is the PURE system?
The PURE system (Protein synthesis Using Recombinant Elements) is a reconstituted cell-free protein synthesis system composed of individually purified components required for transcription and translation.
Unlike conventional cell extract-based systems, the PURE system does not rely on crude cell lysates. Instead, purified components required for protein synthesis are combined in a defined reaction system.
This defined composition provides researchers with a high degree of control over the protein synthesis environment and makes it possible to modify individual components according to experimental needs.
Developed by Professor Takuya Ueda
The PURE system was originally developed by Professor Takuya Ueda and his research group at the University of Tokyo. He currently serves as an advisor to GeneFrontier, bringing his expertise in PURE system technology to the company.
PURE system at a Glance
Defined Composition
A fully reconstituted system built from purified components
Minimal Background Interference
Reduced unwanted enzymatic activities
High Reproducibility
Defined composition supports consistent experimental conditions
Highly Customizable
Precise control of individual reaction components
1. Shimizu et al. (2001) Nat. Biotecnol., vol. 19, p. 751
2. Shimizu et al. (2005) Methods, vol. 36, p. 299
How Does It Work?
The PURE system reconstructs the essential processes required for protein synthesis—including transcription, aminoacylation, energy regeneration, and translation—using purified components.
The diagram below illustrates how these components work together to synthesize a protein from a DNA template.
Components of the PURE system
The PURE system is reconstructed from purified components involved in bacterial transcription and translation.
These include ribosomes, tRNAs, amino acids, aminoacyl-tRNA synthetases, translation factors, RNA polymerase, and components required for energy regeneration.
Translation-related components include initiation factors (IF1, IF2 and IF3), elongation factors (EF-Tu, EF-Ts and EF-G), release factors (RF1, RF2 and RF3), and the ribosome recycling factor (RRF).
Because the reaction is reconstructed from defined components, individual components can be added, removed, or adjusted according to experimental requirements.
Differences from Cell Extract-Based Systems
Both the PURE system and cell extract-based systems, also known as cell lysate-based systems, are widely used for cell-free protein synthesis.
The key difference lies in how the protein synthesis machinery is prepared: the PURE system is reconstituted from individually purified components, whereas extract-based systems use cellular extracts containing the native protein synthesis machinery.
| Comparison | PURE system | Cell Extract-Based System |
|---|---|---|
| System preparation | Reconstituted from individually purified components | Prepared from cellular extracts containing native protein synthesis machinery |
| Composition | Defined set of purified components | Complex mixture of cellular components |
| Background enzymatic activities | Minimized unwanted enzymatic activities with individually purified components | Endogenous enzymatic activities may remain, depending on extract preparation |
| Batch-to-batch variability | Defined composition helps minimize batch-to-batch variability | Batch-to-batch variability may arise from differences in cell culture conditions and extract preparation |
| Component adjustment | Individual components can be added, removed, or adjusted | Reaction conditions can be adjusted, but individual components are less readily controlled |
| Research applications | Suitable for studies requiring precise control of reaction composition | Suitable for applications using the cellular machinery retained in extracts |
These differences make the two approaches suitable for different experimental needs. The PURE system provides a defined reaction environment in which individual components can be controlled, making it particularly useful for studies that require precise manipulation of the protein synthesis machinery.
Research Applications
The PURE system has been used in a variety of research applications, including protein modification, non-canonical amino acid incorporation, and translation studies. Explore selected examples below.
Non-Canonical Amino Acid Incorporation
Amber suppression for the incorporation of non-canonical amino acids into proteins using the PURE system.
N-terminal Modification of Proteins
N-terminal acetylation and myristoylation of proteins synthesized using a cell-free system.
Bring PURE system Technology to Your Research
PUREfrex® is GeneFrontier's reconstituted cell-free protein synthesis system based on PURE system technology.
Explore PUREfrex® Resources
Scientific and Technical Information on PUREfrex®
Tech Notes
Find practical technical information and tips for cell-free protein synthesis.
Cases
Discover examples of protein synthesis and research applications using PUREfrex®.