Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus.
The nonstructural protein VP9 from white spot syndrome virus (WSSV) has been identified and expressed in Escherichia coli. To facilitate purification, a cleavable His6 tag was introduced at the N-terminus. The native protein was purified and crystallized by vapour diffusion against mother liquor containing 2 M sodium acetate, 100 mM MES pH 6.3, 25 mM cadmium sulfate and 3% glycerol. Crystals were obtained within 7 d and diffracted to 2.2 angstroms; they belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 74.13, b = 78.21, c = 78.98 angstroms and four molecules in the asymmetric unit. The selenomethionine-labelled protein produced isomorphous crystals that diffracted to approximately 3.3 angstroms.
Liu
Yang
Y
Department of Biological Sciences, National University of Singapore, Singapore.
Sivaraman
J
J
Hew
Choy L
CL
D003001:Cloning, Molecular; D003460:Crystallization; D017931:DNA Primers; D004926:Escherichia coli; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D016324:Sequence Tagged Sites; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D046848:White spot syndrome virus 1 / Q000737:chemistry* / Q000235:genetics; D014961:X-Ray Diffraction
D017931:DNA Primers; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins