SpringerProtocols: Abstract: 12. Cell-Free Protein Synthesis With Prokaryotic Combined Transcription-Translation

12. Cell-Free Protein Synthesis With Prokaryotic Combined Transcription-Translation

By: James R. Swartz³, Michael C. Jewett³, Kim A. Woodrow³

Abstract

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Cell-free biology exploits and studies complex biological processes in a controlled environment without intact cells. One model system is prokaryotic cell-free protein synthesis. This technology offers an attractive and convenient approach to produce properly folded recombinant DNA (rDNA) proteins on a laboratory scale, screen PCR fragment libraries in a high-throughput format, express pharmaceutical proteins, incorporate labeled or unnatural amino acids into proteins, and activate microbial physiology to allow for investigation of biological systems. We describe the preparation of materials necessary for the expression, quantification, and purification of rDNA proteins from active Escherichia coli extracts.

Affiliation(s): (3) Department of Chemical Engineering, Stanford University, Stanford, CA

Book Title: Recombinant Gene Expression: Reviews and Protocols

Series: Methods in Molecular Biology | Volume: 267 | Pub. Date: Jul-20-2004 | Page Range: 169-182 | DOI: 10.1385/1-59259-774-2:169

Subject: Genetics/Genomics

Key Words: Prokaryotic combined transcription-translation - S30 extract - cell-free protein synthesis - in vitro - chloramphenicol acetyl transferase - T7 bacteriophage RNA polymerase - protein purification

References

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