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Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli

The RNA-dependent-RNA polymerase (RdRp) from SARS-CoV-2 is an important drug target because it is responsible for viral RNA genome replication. Efficient production of recombinant RdRp is important in screening antivirals to treat COVID-19. Here, we present our protocol for expression of tag-free re...

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Autores principales: Dangerfield, Tyler L., Huang, Nathan Z., Johnson, Kenneth A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7859716/
https://www.ncbi.nlm.nih.gov/pubmed/33558863
http://dx.doi.org/10.1016/j.xpro.2021.100357
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author Dangerfield, Tyler L.
Huang, Nathan Z.
Johnson, Kenneth A.
author_facet Dangerfield, Tyler L.
Huang, Nathan Z.
Johnson, Kenneth A.
author_sort Dangerfield, Tyler L.
collection PubMed
description The RNA-dependent-RNA polymerase (RdRp) from SARS-CoV-2 is an important drug target because it is responsible for viral RNA genome replication. Efficient production of recombinant RdRp is important in screening antivirals to treat COVID-19. Here, we present our protocol for expression of tag-free replication complex proteins in E. coli and subsequent purification. Despite the added complexity of multiple purification steps, our methods provide greater activity, yield at lower cost, and are faster than baculovirus expression systems. For complete details on the use and execution of this protocol, please refer to Dangerfield et al. (2020).
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spelling pubmed-78597162021-02-04 Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli Dangerfield, Tyler L. Huang, Nathan Z. Johnson, Kenneth A. STAR Protoc Protocol The RNA-dependent-RNA polymerase (RdRp) from SARS-CoV-2 is an important drug target because it is responsible for viral RNA genome replication. Efficient production of recombinant RdRp is important in screening antivirals to treat COVID-19. Here, we present our protocol for expression of tag-free replication complex proteins in E. coli and subsequent purification. Despite the added complexity of multiple purification steps, our methods provide greater activity, yield at lower cost, and are faster than baculovirus expression systems. For complete details on the use and execution of this protocol, please refer to Dangerfield et al. (2020). Elsevier 2021-02-04 /pmc/articles/PMC7859716/ /pubmed/33558863 http://dx.doi.org/10.1016/j.xpro.2021.100357 Text en © 2021 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Dangerfield, Tyler L.
Huang, Nathan Z.
Johnson, Kenneth A.
Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli
title Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli
title_full Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli
title_fullStr Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli
title_full_unstemmed Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli
title_short Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli
title_sort expression and purification of tag-free sars-cov-2 rna-dependent rna polymerase in escherichia coli
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7859716/
https://www.ncbi.nlm.nih.gov/pubmed/33558863
http://dx.doi.org/10.1016/j.xpro.2021.100357
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