/
/
Cell-Free Protein Synthesis from Exonuclease-Deficient Cellular Extracts Utilizing Linear DNA Templates
A subscription to JoVE is required to view this content. Sign in or start your free trial.
1INRAe, AgroParisTech, Micalis Institute,Université Paris-Saclay, 2Helmholtz Institute for RNA-based Infection Research (HIRI),Helmholtz-Centre for Infection Research (HZI), 3Medical Faculty,University of Würzburg, 4Centre de Biochimie Structurale, INSERM U1054, CNRS UMR 5048,University of Montpellier
Chapters
- 00:05Introduction
- 00:50Cell Culture and Lysate Preparation
- 04:17Cell-Free Buffer Calibration for Linear DNA
- 05:49Experimental Execution
- 06:33Results: Expression in Cell-Free Protein Synthesis Systems Using Linear DNA Templates
- 07:26Conclusion
Presented here is a protocol for the preparation and buffer calibration of cell extracts from exonuclease V knockout strains of Escherichia coli BL21 Rosetta2 (ΔrecBCD and ΔrecB). This is a fast, easy, and direct approach for expression in cell-free protein synthesis systems using linear DNA templates.
