Simple Extract Preparation Methods for E. coli-Based Cell-Free Expression.

Autor:	Mullin AC; Department of Chemistry and Biochemistry, California Polytechnic State University, San Luis Obispo, CA, USA., Slouka T; Department of Chemistry and Biochemistry, California Polytechnic State University, San Luis Obispo, CA, USA., Oza JP; Department of Chemistry and Biochemistry, California Polytechnic State University, San Luis Obispo, CA, USA. joza@calpoly.edu.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2433, pp. 51-64.
DOI:	10.1007/978-1-0716-1998-8_2
Abstrakt:	Cell-free protein synthesis (CFPS) is a powerful platform for synthetic biology, allowing for the controlled expression of proteins without reliance on living cells. However, the process of producing the cell extract, a key component of cell-free reactions, can be a bottleneck for new users to adopt CFPS as it requires technical knowledge and significant researcher oversight. Here, we provide a detailed method for implementing a simplified cell extract preparation workflow using CFAI media. We also provide a detailed protocol for the alternative, 2x YPTG media-based preparation process, as it represents a useful benchmark within the cell-free community. (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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