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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/45976

Title: The unique stacked rings in the nucleocapsid of the WSSV virion are formed by the major structural protein VP664, the largest viral structural protein ever found
Authors: Jiann-Horng Leu;Jyh-Ming Tsai;Han-Ching Wang;Andrew H.-J. Wang;Chung-Hsiung Wang;Guang-Hsiung Kou;Chu-Fang Lo
Contributors: 國立臺灣海洋大學:海洋生物研究所
Date: 2005
Issue Date: 2018-04-18T07:51:00Z
Publisher: Journal of Virology
Abstract: Abstract: One unique feature of the shrimp white spot syndrome virus (WSSV) genome is the presence of a giant open reading frame (ORF) of 18,234 nucleotides that encodes a long polypeptide of 6,077 amino acids with a hitherto unknown function. In the present study, by applying proteomic methodology to analyze the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile of purified WSSV virions by liquid chromatography-mass spectrometry (LC-MS/MS), we found that this giant polypeptide, designated VP664, is one of the viral structural proteins. The existence of the corresponding 18-kb transcript was confirmed by sequencing analysis of reverse transcription-PCR products, which also showed that vp664 was intron-less. A time course analysis showed that this transcript was actively transcribed at the late stage, suggesting that this gene product should contribute primarily to the assembly and morphogenesis of the virion. Several polyclonal antisera against this giant protein were prepared, and one of them was successfully used for immunoelectron microscopy analysis to localize the protein in the virion. Immunoelectron microscopy with a gold-labeled secondary antibody showed that the gold particles were regularly distributed around the periphery of the nucleocapsid with a periodicity that matched the characteristic stacked ring subunits that appear as striations. From this and other evidence, we argue that this giant ORF in fact encodes the major WSSV nucleocapsid protein.
Relation: 79(1) pp.140–149
URI: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/45976
Appears in Collections:[海洋生物研究所] 期刊論文

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