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Published on: April 19, 2016
Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus
Yang Liu1, J Sivaraman, Choy L Hew
1Department of Biological Sciences, National University of Singapore, Singapore.
Abstract:
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.

