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Simple procedures for the construction of a robust and cost-effective cell-free protein synthesis system

Paper ID Volume ID Publish Year Pages File Format Full-Text
25625 43585 2006 8 PDF Available
Title
Simple procedures for the construction of a robust and cost-effective cell-free protein synthesis system
Abstract

In this study, as a part of our efforts to improve the robustness and economical feasibility of cell-free protein synthesis, we developed a simple method of preparing the cell extracts used for catalyzing cell-free protein synthesis reactions. We found that the high-speed centrifugation, pre-incubation, and dialysis steps of the conventional procedures could be omitted without losing the translational activity of the resulting cell extract. Instead, a simple centrifugation step at low speed (12,000 RCF for 10 min) followed by a brief period of incubation was sufficient for the preparation of an active extract to support cell-free protein synthesis with higher productivity and consistency. Compared to the present standard procedures for the preparation of the S30 extract, the overall cost of the reagents and processing time were reduced by 80 and 60%, respectively.

Keywords
S30 extract; Cell-free protein synthesis; Escherichia coli; Pre-incubation; Chloramphenicol acetyltransferase (CAT)
First Page Preview
Simple procedures for the construction of a robust and cost-effective cell-free protein synthesis system
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 126, Issue 4, 1 December 2006, Pages 554–561
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering