Elimination of ribosome inactivating factors improves the efficiency of and cell-free translation systems

Brodiazhenko, Tetiana, Johansson, Marcus J O, Takada, Hiraku, Nissan, Tracy, Hauryliuk, Vasili and Murina, Victoriia (2018) Elimination of ribosome inactivating factors improves the efficiency of and cell-free translation systems. Frontiers in Microbiology, 9 (3041). pp. 1-9. ISSN 1664-302X

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Abstract

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.

Item Type: Article
Schools and Departments: School of Life Sciences > Biochemistry
Subjects: Q Science > QR Microbiology > QR0001 General
Q Science > QR Microbiology > QR0075 Bacteria
Depositing User: Tracy Nissan
Date Deposited: 15 Jul 2019 08:27
Last Modified: 08 Oct 2019 07:01
URI: http://sro.sussex.ac.uk/id/eprint/84898

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