Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of the structural domain of the nucleocapsid N protein from porcine reproductive and respiratory syndrome virus (PRRSV).
The structural domain of the PRRSV nucleocapsid N protein was overexpressed in Escherichia coli and purified to homogeneity. Crystals of the expressed protein, designated His-Ndelta(57), were obtained by hanging-drop vapour diffusion using PEG 3350 as precipitant at pH 6.5. A native data set from a frozen crystal was collected to 2.7 A resolution using synchrotron radiation. The crystals belong to space group P3(1)21 or P3(2)21, with unit-cell parameters a = 44.41, c = 125.05, and contain a dimer in the asymmetric unit.
Doan
Danny N P
DN
Institute of Molecular and Cell Biology, 30 Medical Drive, Singapore 117609, Singapore. doan@imcb.nus.edu.sg
Dokland
Terje
T
D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D018076:DNA, Complementary / Q000378:metabolism; D019281:Dimerization; D004926:Escherichia coli / Q000378:metabolism; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D019590:Nucleocapsid Proteins / Q000737:chemistry*; D019316:Porcine respiratory and reproductive syndrome virus / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D014961:X-Ray Diffraction
D018076:DNA, Complementary; D019590:Nucleocapsid Proteins; D006639:Histidine