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Encompassing Amino Acids: NSP8_NSP7: 1-242(end); NSP12: 1-932(end)
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: RNA-dependent RNA polymerase was incubated with FRET Dig-RNA Duplex or Eu-RNA Duplex in the presence of RNaseOUT and biotinylated ATP for 60 min at 37°C in a 10-µl reaction. Streptavidin-acceptor dye and Eu-labeled a-Dig antibody (for Dig-labeled substrate) was then added followed by TR-FRET detection (317 => 620 nm, and 317 => 665 nm). Increase in TR-FRET is proportional to the amount of ATP incorporated in the RNA.
Description: Recombinant complex of three SARS-CoV-2 proteins NSP7 (nonstructural protein 7), NSP8 (nonstructural protein 8) and NSP12 (nonstructural protein 12). The NSP8_NSP7 construct contains an N-terminal His-tag (14xHis) followed by a TEV protease target sequence that allows cleavage of His-tag (TEV: tobacco etch virus cysteine protease BPS Bioscience #50308). The recombinant NSP12 construct is full length encompassing amino acids 1-932 (end). These recombinant proteins are co-expressed in an E. coli expression system and affinity purified.
Format: Aqueous buffer solution
Formulation: 20 mM HEPES, pH 8.0, 300 mM NaCl, 10% glycerol, and 3 mM DTT
Genbank: NSP7: YP_009742614; NSP8: YP 009725304.1; NSP12: YP_009725307
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Tags: NSP8_NSP7: N-terminal His-tag
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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