Expression and purification of tag-free SARS-CoV-2 RNA-dependent RNA polymerase in Escherichia coli.
STAR Protoc
; 2(1): 100357, 2021 03 19.
Article
in English
| MEDLINE | ID: covidwho-1062654
ABSTRACT
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).
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Recombinant Proteins
/
Protein Engineering
/
Escherichia coli
/
Coronavirus RNA-Dependent RNA Polymerase
Limits:
Humans
Language:
English
Journal:
STAR Protoc
Year:
2021
Document Type:
Article
Affiliation country:
J.xpro.2021.100357
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