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

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posted on 2023-06-09, 18:24 authored by Tetiana Brodiazhenko, Marcus J O Johansson, Hiraku Takada, Tracy Nissan, Vasili Hauryliuk, Victoriia Murina
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.

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Publication status

  • Published

File Version

  • Published version

Journal

Frontiers in Microbiology

ISSN

1664-302X

Publisher

Frontiers Media

Issue

3041

Volume

9

Page range

1-9

Department affiliated with

  • Biochemistry Publications

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2019-07-15

First Open Access (FOA) Date

2019-07-15

First Compliant Deposit (FCD) Date

2019-07-15

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