STAR Protocols
Volume 2, Issue 1, 19 March 2021, 100357
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Protocol
Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli

https://doi.org/10.1016/j.xpro.2021.100357Get rights and content
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Highlights

  • Soluble expression of tag-free, highly active SARS-CoV-2 polymerase complex in E. coli

  • Purification protocol for NSP-7/8/12 RNA polymerase complex is provided

  • Protocol for expression and purification of supplemental NSP8 included

  • Purified SARS-CoV-2 RNA polymerase complex catalyzes fast processive RNA replication

Summary

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).

Subject areas

Artificial intelligence
Human-centered computing
Human-computer interaction

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