In:
RNA, Cold Spring Harbor Laboratory, Vol. 27, No. 11 ( 2021-11), p. 1390-1399
Abstract:
In bacteria, trans -translation is the major quality control system for rescuing stalled ribosomes. It is mediated by tmRNA, a hybrid RNA with properties of both a tRNA and a mRNA, and the small protein SmpB. Because trans -translation is absent in eukaryotes but necessary for bacterial fitness or survival, it is a promising target for the development of novel antibiotics. To facilitate screening of chemical libraries, various reliable in vitro and in vivo systems have been created for assessing trans -translational activity. However, the aim of the current work was to permit the safe and easy in vitro evaluation of trans -translation from pathogenic bacteria, which are obviously the ones we should be targeting. Based on green fluorescent protein (GFP) reassembly during active trans -translation, we have created a cell-free assay adapted to the rapid evaluation of trans -translation in ESKAPE bacteria, with 24 different possible combinations. It can be used for easy high-throughput screening of chemical compounds as well as for exploring the mechanism of trans -translation in these pathogens.
Type of Medium:
Online Resource
ISSN:
1355-8382
,
1469-9001
DOI:
10.1261/rna.078773.121
Language:
English
Publisher:
Cold Spring Harbor Laboratory
Publication Date:
2021
detail.hit.zdb_id:
1475737-0
SSG:
12
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