Expression and purification of the full-length SARS-CoV-2 nucleoprotein (IMAGE)
Caption
(A) Map of the pHYP-NPC-10H plasmid. 10xHIS—C-terminal decahistidine tag; KanR2—kanamycin A resistance gene; HS—hok/sok post—segregation killing locus; lac I—gene of the LacI repressor protein; T7 prom—promoter for bacteriophage T7 RNA polymerase. (B) SDS-PAGE analysis of the full—length NP expression. Induction with 1 mM IPTG. (C) SDS-PAGE analysis of the NP purification in the presence or absence of the RNase A. Target protein position is marked with the red arrow. “Non-binding”—column flowthrough fraction. (D, E) IMAC chromatography traces for the NP purification in the presence or absence of RNase A treatment and 2 M NaCl solution wash. Eluate absorbance at 280 nm is in blue, absorbance at 260 nm—in red. (F) Agarose gel analysis of purified proteins, ethidium bromide staining, double-strand DNA marker.
Credit
Kolesov DE, Sinegubova MV, Safenkova IV, Vorobiev II, Orlova NA. 2022. Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture. PeerJ
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License
CC BY-NC