Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus.
Abstract
The nonstructural protein VP9 from white spot syndrome virus (WSSV) has been identified and expressed in Escherichia coli. To facilitate purification, a cleavable His{sub 6} tag was introduced at the N-terminus. The native protein was purified and crystallized by vapor 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{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 74.13, b = 78.21, c = 78.98 Angstroms and four molecules in the asymmetric unit. The selenomethionine-labeled protein produced isomorphous crystals that diffracted to approximately 3.3 Angstroms.
- Authors:
- Publication Date:
- Research Org.:
- Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source
- Sponsoring Org.:
- Doe - Office Of Science
- OSTI Identifier:
- 914148
- Report Number(s):
- BNL-78716-2007-JA
TRN: US0801575
- DOE Contract Number:
- DE-AC02-98CH10886
- Resource Type:
- Journal Article
- Journal Name:
- Acta Cryst. F
- Additional Journal Information:
- Journal Volume: 62; Journal Issue: 8
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 43 PARTICLE ACCELERATORS; CADMIUM SULFATES; CRYSTALLIZATION; DIFFUSION; ESCHERICHIA COLI; GLYCEROL; PROTEINS; PURIFICATION; SODIUM; SPACE GROUPS; NSLS; national synchrotron light source
Citation Formats
Liu, Y, Sivaraman, J, and Hew, C. Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus.. United States: N. p., 2006.
Web. doi:10.1107/S1744309106027436.
Liu, Y, Sivaraman, J, & Hew, C. Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus.. United States. https://doi.org/10.1107/S1744309106027436
Liu, Y, Sivaraman, J, and Hew, C. 2006.
"Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus.". United States. https://doi.org/10.1107/S1744309106027436.
@article{osti_914148,
title = {Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus.},
author = {Liu, Y and Sivaraman, J and Hew, C},
abstractNote = {The nonstructural protein VP9 from white spot syndrome virus (WSSV) has been identified and expressed in Escherichia coli. To facilitate purification, a cleavable His{sub 6} tag was introduced at the N-terminus. The native protein was purified and crystallized by vapor 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{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 74.13, b = 78.21, c = 78.98 Angstroms and four molecules in the asymmetric unit. The selenomethionine-labeled protein produced isomorphous crystals that diffracted to approximately 3.3 Angstroms.},
doi = {10.1107/S1744309106027436},
url = {https://www.osti.gov/biblio/914148},
journal = {Acta Cryst. F},
number = 8,
volume = 62,
place = {United States},
year = {Sun Jan 01 00:00:00 EST 2006},
month = {Sun Jan 01 00:00:00 EST 2006}
}