Effects of growth rate on cell extract performance in cell-free protein synthesis

2006 ◽  
Vol 94 (4) ◽  
pp. 618-624 ◽  
Author(s):  
James Zawada ◽  
James Swartz
Keyword(s):  
Growth Rate ◽  
Protein Synthesis ◽  
Cell Extract ◽  
Free Protein ◽  
Cell Free Protein Synthesis

Efficiency of cell-free protein synthesis based on a crude cell extract from Escherichia coli, wheat germ, and rabbit reticulocytes

2008 ◽  
Vol 133 (2) ◽  
pp. 183-189 ◽  
Author(s):  
Mami Hino ◽  
Masatoshi Kataoka ◽  
Kazuaki Kajimoto ◽  
Takenori Yamamoto ◽  
Jun-Ichi Kido ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein Synthesis ◽  
Wheat Germ ◽  
Cell Extract ◽  
Crude Cell Extract ◽  
Free Protein ◽  
Cell Free Protein Synthesis ◽  
Rabbit Reticulocytes

scholarly journals A Crude Extract Preparation and Optimization from a Genomically Engineered Escherichia coli for the Cell-Free Protein Synthesis System: Practical Laboratory Guideline

2019 ◽  
Vol 2 (3) ◽  
pp. 68 ◽  
Author(s):  
Kim ◽  
Copeland ◽  
Padumane ◽  
Kwon
Keyword(s):  
Escherichia Coli ◽  
Protein Synthesis ◽  
Optimization Procedure ◽  
Cell Extract ◽  
Coli Cell ◽  
Free Protein ◽  
E Coli ◽  
Highly Active ◽  
Cell Free Protein Synthesis

With the advancement of synthetic biology, the cell-free protein synthesis (CFPS) system has been receiving the spotlight as a versatile toolkit for engineering natural and unnatural biological systems. The CFPS system reassembles the materials necessary for transcription and translation and recreates the in vitro protein synthesis environment by escaping a physical living boundary. The cell extract plays an essential role in this in vitro format. Here, we propose a practical protocol and method for Escherichia coli-derived cell extract preparation and optimization, which can be easily applied to both commercially available and genomically engineered E. coli strains. The protocol includes: (1) The preparation step for cell growth and harvest, (2) the thorough step-by-step procedures for E. coli cell extract preparation including the cell wash and lysis, centrifugation, runoff reaction, and dialysis, (3) the preparation for the CFPS reaction components and, (4) the quantification of cell extract and cell-free synthesized protein. We anticipate that the protocol in this research will provide a simple preparation and optimization procedure of a highly active E. coli cell extract.

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