Production of coronavirus nonstructural proteins in soluble form for crystallization.
For biophysical investigations on viral proteins, in particular for structure determination by X-ray crystallography, relatively large quantities of purified protein are necessary. However, expression of cDNAs coding for viral proteins in prokaryotic or eukaryotic systems is often not straightforward, and frequently the amount and/or the solubility of the protein obtained are not sufficient. Here, we describe a number of protocols for production of nonstructural proteins of coronaviruses that have proven to be efficient in increasing expression yields or solubilities.
Piotrowski
Yvonne
Y
Center for Structural and Cell Biology in Medicine, Institute of Biochemistry, University of Lübeck, Germany.
Ponnusamy
Rajesh
R
Glaser
Stephanie
S
Daabach
Anniken
A
Moll
Ralf
R
Hilgenfeld
Rolf
R
D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012995:Solubility; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*
D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C087633:nonstructural protein, coronavirus