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

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 ta...

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Bibliographic Details
Main Authors: Tyler L. Dangerfield, Nathan Z. Huang, Kenneth A. Johnson
Format: Article
Language:English
Published: Elsevier 2021-03-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721000642
Description
Summary: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).
ISSN:2666-1667