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Crystallization and X‐ray diffraction of virus‐like particles from a piscine betanodavirus
Author(s) -
Luo YuChun,
Wang ChunHsiung,
Wu YiMin,
Liu Wangta,
Lu MingWei,
Lin ChanShing
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14013703
Subject(s) - capsid , grouper , virus , biology , crystallography , open reading frame , virus like particle , virology , chemistry , peptide sequence , fish <actinopterygii> , gene , recombinant dna , genetics , fishery
Dragon grouper nervous necrosis virus (DGNNV), a member of the genus Betanodavirus , causes high mortality of larvae and juveniles of the grouper fish Epinephelus lanceolatus . Currently, there is no reported crystal structure of a fish nodavirus. The DGNNV virion capsid is derived from a single open reading frame that encodes a 338‐amino‐acid protein of approximately 37 kDa. The capsid protein of DGNNV was expressed to form virus‐like particles (VLPs) in Escherichia coli . The VLP shape is T = 3 quasi‐symmetric with a diameter of ∼38 nm in cryo‐electron microscopy images and is highly similar to the native virion. In this report, crystals of DGNNV VLPs were grown to a size of 0.27 mm within two weeks by the hanging‐drop vapour‐diffusion method at 283 K and diffracted X‐rays to ∼7.5 Å resolution. In‐house X‐ray diffraction data of the DGNNV VLP crystals showed that the crystals belonged to space group R 32, with unit‐cell parameters a = b = 353.00, c = 800.40 Å, α = β = 90, γ = 120°. 23 268 unique reflections were acquired with an overall R merge of 18.2% and a completeness of 93.2%. Self‐rotation function maps confirmed the fivefold, threefold and twofold symmetries of the icosahedron of DGNNV VLPs.