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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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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). |
format | Online Article Text |
id | pubmed-7859716 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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