California Polytechnic State University San Luis Obispo
2 ARTICLES PUBLISHED IN JoVE
Chemistry
Escherichia coli-Based Cell-Free Protein Synthesis: Protocols for a robust, flexible, and accessible platform technology
Max Z. Levine *1,2, Nicole E. Gregorio *2,3, Michael C. Jewett 4,5,6,7, Katharine R. Watts 2,3, Javin P. Oza 2,3
1Department of Biological Sciences, California Polytechnic State University, San Luis Obispo, 2Center for Applications in Biotechnology, California Polytechnic State University, San Luis Obispo, 3Department of Chemistry and Biochemistry, California Polytechnic State University, 4Department of Chemical and Biological Engineering, Northwestern University, 5Chemistry of Life Processes Institute, Northwestern University, 6Center for Synthetic Biology, Northwestern University, 7Interdisciplinary Biological Sciences Program, Northwestern University
This protocol details the steps, costs, and equipment necessary to generate E. coli-based cell extracts and implement in vitro protein synthesis reactions within 4 days or less. To leverage the flexible nature of this platform for broad applications, we discuss reaction conditions that can be adapted and optimized.
Bioengineering
From Cells to Cell-Free Protein Synthesis within 24 Hours Using Cell-Free Autoinduction Workflow
Philip E. J. Smith 1,2, Taylor Slouka 1,2, Mona Dabbas 1,2, Javin P. Oza 1,2
1Department of Chemistry and Biochemistry, California Polytechnic State University San Luis Obispo, 2Center for Application in Biotechnology, California Polytechnic State University San Luis Obispo
This work describes the preparation of cell extract from Escherichia coli (E. coli) followed by cell-free protein synthesis (CFPS) reactions in under 24 hours. Explanation of the cell-free autoinduction (CFAI) protocol details improvements made to reduce researcher oversight and increase quantities of cell extract obtained.
