Cell-free protein synthesis from non-growing, stressed Escherichia coli.

ABSTRACT: Cell-free protein synthesis is a versatile protein production system. Performance of the protein synthesis depends on highly active cytoplasmic extracts. Extracts from E. coli are believed to work best; they are routinely obtained from exponential growing cells, aiming to capture the most active translation system. Here, we report an active cell-free protein synthesis system derived from cells harvested at non-growth, stressed conditions. We found a downshift of ribosomes and proteins. However, a characterization revealed that the stoichiometry of ribosomes and key translation factors was conserved, pointing to a fully intact translation system. This was emphasized by synthesis rates, which were comparable to those of systems obtained from fast-growing cells. Our approach is less laborious

SUBMITTER: Failmezger J 

PROVIDER: S-EPMC5705671 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature