Reassembling green fluorescent protein for in vitro evaluation of trans-translation

In order to discover new antibiotics with improved activity and selectivity, we created a reliable in vitro reporter system to detect trans-translation activity, the main mechanism for recycling ribosomes stalled on problematic messenger RNA (mRNA) in bacteria. This system is based on an engineered...

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Detalles Bibliográficos
Autores principales: Guyomar, Charlotte, Thépaut, Marion, Nonin-Lecomte, Sylvie, Méreau, Agnès, Goude, Renan, Gillet, Reynald
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7038980/
https://www.ncbi.nlm.nih.gov/pubmed/31919515
http://dx.doi.org/10.1093/nar/gkz1204
Descripción
Sumario:In order to discover new antibiotics with improved activity and selectivity, we created a reliable in vitro reporter system to detect trans-translation activity, the main mechanism for recycling ribosomes stalled on problematic messenger RNA (mRNA) in bacteria. This system is based on an engineered tmRNA variant that reassembles the green fluorescent protein (GFP) when trans-translation is active. Our system is adapted for high-throughput screening of chemical compounds by fluorescence.

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