Elimination of Ribosome Inactivating Factors Improves the Efficiency of Bacillus subtilis and Saccharomyces cerevisiae Cell-Free Translation Systems

Cell-free translation systems based on cellular lysates optimized for protein synthesis have multiple applications both in basic and applied science, ranging from studies of translational regulation to cell-free production of proteins and ribosome-nascent chain complexes. In order to achieve both hi...

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Bibliographic Details
Published in:Frontiers in microbiology 2018, Vol.9, p.3041-3041
Main Authors: Brodiazhenko, Tetiana, Johansson, Marcus J O, Takada, Hiraku, Nissan, Tracy, Hauryliuk, Vasili, Murina, Victoriia
Format: Article
Language:English
Subjects:
Bacillus subtilis
cell-free translation system
cell-tree translation system
HPF
Microbiology
Saccharomyces cerevisiae
Stm1
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Summary:Cell-free translation systems based on cellular lysates optimized for protein synthesis have multiple applications both in basic and applied science, ranging from studies of translational regulation to cell-free production of proteins and ribosome-nascent chain complexes. In order to achieve both high activity and reproducibility in a translation system, it is essential that the ribosomes in the cellular lysate are enzymatically active. Here we demonstrate that genomic disruption of genes encoding ribosome inactivating factors - HPF in and Stm1 in - robustly improve the activities of bacterial and yeast translation systems. Importantly, the elimination of HPF results in a complete loss of 100S ribosomes, which otherwise interfere with disome-based approaches for preparation of stalled ribosomal complexes for cryo-electron microscopy studies.
ISSN:1664-302X
1664-302X
DOI:10.3389/fmicb.2018.03041