NP - Expression / Purification

Constructs obtained from Geneart/Ralf

-- cloned into pET21a

-- Expression in Rosetta

MERS-CoV-N, HCoV-NL63-N, HCoV-229E-N, HCoV-HKU1-N, HCoV-OC43-N, SARS-CoV-1-N

fehlt noch: SARS-CoV-2-N

Studies show aggregation of fullength NP

(2020 pmid: 32363136) -- full-length SARS-CoV-2 N protein aggregated status were found in our expression and purification studies, as well as previously reported data on other coronavirus nucleocapsid protein.

- we should do a buffer screen - check solubility - this would be great already

Full length expression achieved

(Zheng-2020 doi: 10.1016/j.bbrc.2020.04.136)

(Carlson MolCell 2020 PMID: 33248025) denaturating purification and refolding of fulllength NP - removal of associated RNAs