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Related Concept Videos

Ribosomes01:27

Ribosomes

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Ribosomes translate genetic information encoded by messenger RNA (mRNA) into proteins. Both prokaryotic and eukaryotic cells have ribosomes. Cells that synthesize large quantities of protein—such as secretory cells in the human pancreas—can contain millions of ribosomes.
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Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
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Updated: Apr 23, 2026

Author Spotlight: Optimizing CFPS Systems for Synthetic Cell Construction
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Author Spotlight: Optimizing CFPS Systems for Synthetic Cell Construction

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Protein synthesis using a reconstituted cell-free system.

Corinna Tuckey1, Haruichi Asahara1, Ying Zhou1

  • 1New England Biolabs, Inc, Ipswich, Massachusetts.

Current Protocols in Molecular Biology
|October 2, 2014
PubMed
Summary
This summary is machine-generated.

This study introduces PURExpress, a reconstituted cell-free protein synthesis system. It enables efficient protein production and modification with minimal background activity, offering a purified and adaptable tool for molecular biology research.

Keywords:
PURExpresscoupled transcription and translationisotope labelingreconstituted cell-free protein synthesisribosome displayunnatural amino acid incorporation

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Synthetic Biology

Background:

  • Traditional cell-free protein synthesis (CFPS) systems often utilize crude cell extracts.
  • These extracts can contain nucleases and proteases, limiting protein yield and purity.
  • Reconstituted systems offer a controlled environment using purified components.

Purpose of the Study:

  • To describe the application and advantages of the PURExpress commercial reconstituted CFPS system.
  • To highlight its utility for in vitro protein synthesis, labeling, and purification.
  • To showcase the system's adaptability for specialized applications.

Main Methods:

  • Utilizing the PURExpress system for in vitro protein synthesis from various DNA and mRNA templates.
  • Employing His-tagged components for purification of synthesized proteins.
  • Modifying the system by omitting specific components (e.g., ribosomes, release factors) to create 'delta' versions.

Main Results:

  • PURExpress facilitates efficient protein synthesis with minimal nuclease and protease activity.
  • His-tagged system components allow for straightforward purification of synthesized proteins.
  • Synthesized proteins are often detectable by SDS-PAGE and can be directly functionalized.
  • Modified 'delta' systems enable specific applications like unnatural amino acid incorporation and ribosome display.

Conclusions:

  • The PURExpress system provides a robust and versatile platform for cell-free protein synthesis.
  • Its defined composition and purification capabilities offer advantages over traditional extract-based systems.
  • The system's modularity supports advanced applications in translation studies and protein engineering.