To express the SARS-CoV-2 replicase proteins (NC_045512.2 ), which is made up of nsp1, nsp2, nsp3, nsp5, nsp7, nsp8, nsp9, nsp10, nsp13, nsp14, and nsp15, gene fragment were cloned into the prokaryotic expression vector pET-30a with an N/C-terminal His-tag, yielding plasmids pET-30a-SARS-CoV-2-nsps. These plasmids were transformed into Escherichia coli BL21(DE3) cells, and His-tagged proteins were purified using Ni-NTA His-Tagged (TransGen Biotech) according to the manufacturer’s protocol. Similarly, replicase gene fragments were also cloned into the eukaryotic expression vector pRK5 with a C-terminal Myc tag (pRK5-SARS-CoV-2-nsps) for detection by antisera. Similarly, replicase gene fragments were also cloned into the eukaryotic expression vector pRK5 with an N/C-terminal Myc or Flag tag (pRK5-SARS-CoV-2-nsps) for detection by antisera.
Ni nta his tagged
Manufactured by Transgene
Ni-NTA His-Tagged is a laboratory equipment product designed for the purification of recombinant proteins with a histidine tag. It utilizes a nickel-nitrilotriacetic acid (Ni-NTA) resin to bind and capture the histidine-tagged proteins from complex mixtures.
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Lab products found in correlation
2 protocols using ni nta his tagged
1
Cloning and Purification of SARS-CoV-2 Replicase Proteins
2
Cloning and Purification of SARS-CoV-2 Replicase Proteins
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To express the SARS-CoV-2 replicase proteins (NC_045512.2 ), which is made up of nsp1, nsp2, nsp3, nsp5, nsp7, nsp8, nsp9, nsp10, nsp13, nsp14, and nsp15, gene fragment were cloned into the prokaryotic expression vector pET-30a with an N/C-terminal His-tag, yielding plasmids pET-30a-SARS-CoV-2-nsps. These plasmids were transformed into Escherichia coli BL21(DE3) cells, and His-tagged proteins were purified using Ni-NTA His-Tagged (TransGen Biotech) according to the manufacturer’s protocol. Similarly, replicase gene fragments were also cloned into the eukaryotic expression vector pRK5 with a C-terminal Myc tag (pRK5-SARS-CoV-2-nsps) for detection by antisera. Similarly, replicase gene fragments were also cloned into the eukaryotic expression vector pRK5 with an N/C-terminal Myc or Flag tag (pRK5-SARS-CoV-2-nsps) for detection by antisera.
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