Recombinant 2019nCoV antigen-coronavirus
RNA-directed RNA polymerase(RdRP, Nsp12, His Tag)

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Cat No.: GMP-V-2019nCoV-RdRP001

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1mg

GMP-V-2019nCoV-RdRP001-1mg

 5990
10mg

GMP-V-2019nCoV-RdRP001-10mg

 38790
100mg

GMP-V-2019nCoV-RdRP001-100mg

 258000
≥100mg

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Description

Accession Number QHD43415.1
Expression platform 2019-nCoV(SARS-CoV-2, SARS2-coronavirus)
IsotypesE.coli
TagN-His
Products description Recombinant 2019-nCoV(SARS-CoV-2)RNA-directed RNA polymerase (RdRP, Nsp12) was expressed in
E.coli-based prokaryotic cell expression system and the target gene encoding Asp4891-Val5212
was expressed with His tag at the N-terminus.
Bioactivity validationIn Testing
PurityPurity: ≥95% (SDS-PAGE)
ApplicationImmunogen in Elisa,lateral-flow tests,and other immunoassays;Standard substance
The antigen can also be used in drug discovery including antibody screening and lead compound candidates assay.
predicted Molecular Mass39.4kD
Formulation & ReconstitutionLyophilized from GM's Protein Stability Buffer2 (PSB2,Confidential Ingredients) or PBS (pH7.4);
For PSB2, reconstituted with 0.9% sodium chloride;
For PBS, reconstituted with ddH2O.
StorageStore at -20℃ to -80℃ under sterile conditions. Avoid repeated freeze-thaw cycles.

2019nCoV (SARS-CoV-2) RNA-Dependent RNA Polymerase (RdRP, Nsp12)


About Coronavirus RNA-Dependent RNA Polymerase (Coronavirus RdRP)

COVID-19 pandemic is caused by 2019nCoV (SARS-CoV-2) infection. 2019nCoV contains 16 Non-structure protein (Nsp1-Nsp16) that may be drugable targets for antiviral compounds discovery against COVID-191.

2019nCoV (SARS-CoV-2) RNA-Dependent RNA Polymerase (RdRP, Nsp12) is a multidomain RNA polymerase, which is the most conserved protein in coronaviruses. The coronavirus RNA-dependent RNA polymerase (RdRp) is essential for the viral replication and transcription.1

High-throughput screening (HTS) assay development of Coronavirus RdRP for antiviral compounds

HTS Approach: Fluorescence based assay for time-dependent kinetic analysis

Assay Substrate

1.γ-(BBT)-ATP (γ-[2'(2-benzothiazoyl)-6'-hydroxybenzothiazole)-adenosine-5'-triphosphate, Sodium salt)2,3
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Figure. Structural formula of γ-(BBT)-ATP

2. Self-priming RNA (fluorometric)2 OR 3′UTR-U30 RNA (radioactive)3

Simple 2019nCoV (SARS-CoV-2) RNA-Dependent RNA Polymerase (RdRP) activity assay develoement Protocol2,3:
2019nCoV (SARS-CoV-2) RNA-Dependent RNA Polymerase (RdRP)activity assay develoement Protocol-radioactive pdf download
2019nCoV (SARS-CoV-2) RNA-Dependent RNA Polymerase (RdRP)activity assay develoement Protocol-self priming pdf download

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Figure. Schematic representative of Self-priming RNA-based Fluorometric RdRp activity assay


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Figure: Screening workflow of the representative library in 384-well plates. Cip, calf intestinal alkaline phosphatase; RT, room temperature2,3.

References

1. Gordon, D. E. et al. A SARS-CoV-2-Human Protein-Protein Interaction Map Reveals Drug Targets and Potential Drug-Repurposing. BioRxiv, doi:10.1101/2020.03.22.002386 (2020).
2. Kocabas, F., Turan, R. D. & Aslan, G. S. Fluorometric RdRp assay with self-priming RNA. Virus Genes 50, 498-504, doi:10.1007/s11262-015-1187-8 (2015).
3. Niyomrattanakit, P. et al. A fluorescence-based alkaline phosphatase-coupled polymerase assay for identification of inhibitors of dengue virus RNA-dependent RNA polymerase. J Biomol Screen 16, 201-210, doi:10.1177/1087057110389323 (2011).