Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Video

Updated: Jul 11, 2026

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
11:19

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses

Published on: February 26, 2011

Cell-free translation reconstituted with purified components.

Y Shimizu1, A Inoue, Y Tomari

  • 1Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Bldg FSB401, 5-1-5, Kasiwanoha, Kashiwa, Chiba prefecture 277-8562, Japan.

Nature Biotechnology
|August 2, 2001
PubMed
Summary

Researchers created a cell-free protein synthesis system for efficient protein production and purification. This system also enables the incorporation of unnatural amino acids, advancing synthetic biology applications.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

IL-6 in a pleomorphic type of malignant fibrous histiocytoma presenting high fever.

Human pathology·1998
Same author

The effect of pentoxifylline (PTX) on Theiler's murine encephalomyelitis (TMEV)-induced demyelinating disease.

Cellular immunology·1998
Same author

Biomechanical effect and clinical application of the hip joint moment reduction brace.

Clinical orthopaedics and related research·1998
Same author

Differential effects on D2 dopamine receptor and prolactin gene expression by haloperidol and aripiprazole in the rat pituitary.

Brain research. Molecular brain research·1998
Same author

Modified Chiari osteotomy for arthrosis after Perthes' disease. 14 hips followed for 2-12 years.

Acta orthopaedica Scandinavica·1998
Same author

Endothelins inhibit mineralization of rat calvarial osteoblast-like cells.

Journal of cardiovascular pharmacology·1998

Area of Science:

  • Biochemistry
  • Synthetic Biology
  • Molecular Biology

Background:

  • Cell-free protein synthesis (CFPS) systems offer a powerful platform for producing proteins.
  • Existing CFPS systems often require complex optimization and purification protocols.
  • Expanding the capabilities of CFPS to include non-natural amino acid incorporation is a key goal.

Purpose of the Study:

  • To develop a reconstituted, cell-free protein synthesis system using purified components.
  • To achieve high protein production yields and facilitate straightforward purification.
  • To demonstrate the system's capacity for incorporating unnatural amino acids.

Main Methods:

  • Reconstitution of a protein-synthesizing system from purified, tagged recombinant protein factors.

More Related Videos

Functional Surface-immobilization of Genes Using Multistep Strand Displacement Lithography
11:05

Functional Surface-immobilization of Genes Using Multistep Strand Displacement Lithography

Published on: October 25, 2018

Cell-Free Production of Proteoliposomes for Functional Analysis and Antibody Development Targeting Membrane Proteins
08:46

Cell-Free Production of Proteoliposomes for Functional Analysis and Antibody Development Targeting Membrane Proteins

Published on: September 22, 2020

Related Experiment Videos

Last Updated: Jul 11, 2026

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
11:19

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses

Published on: February 26, 2011

Functional Surface-immobilization of Genes Using Multistep Strand Displacement Lithography
11:05

Functional Surface-immobilization of Genes Using Multistep Strand Displacement Lithography

Published on: October 25, 2018

Cell-Free Production of Proteoliposomes for Functional Analysis and Antibody Development Targeting Membrane Proteins
08:46

Cell-Free Production of Proteoliposomes for Functional Analysis and Antibody Development Targeting Membrane Proteins

Published on: September 22, 2020

  • Batch mode protein production assays.
  • Affinity chromatography for purification of tagged protein products.
  • Omission of release factor and addition of suppressor transfer RNA (tRNA) for unnatural amino acid incorporation.

    • Main Results:

    • The reconstituted system achieved a protein production rate of approximately 160 microg/ml/h in batch mode.
    • Protein products were efficiently purified within 1 hour using affinity chromatography.
    • The system successfully facilitated the incorporation of an unnatural amino acid when a release factor was omitted.

    Conclusions:

    • A highly efficient and easily purified cell-free protein synthesis system has been developed.
    • The system's ability to incorporate unnatural amino acids broadens its potential applications in protein engineering and synthetic biology.
    • This reconstituted system provides a streamlined approach for producing and modifying proteins in vitro.