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Submitted on December 14, 2001
Department of Biological Sciences, National University of Singapore, 117543
Corresponding Author: dbshead{at}nus.edu.sg
White spot syndrome virus (WSSV) is at present, one of the major pathogens in shrimp culture worldwide. The complete genome of this virus has recently been sequenced. To identify the structural and functional proteins of WSSV, the purified virions were separated by SDS-PAGE. Twenty-four protein bands were excised, in-gel digested with trypsin and subjected to matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS), respectively. 18 proteins matching the open reading frames (ORFs) of WSSV genome were identified. Except for three known structural proteins and collagen, the functions of the remaining 14 proteins were unknown. Temporal analysis revealed that all the genes were transcribed in the late stage of WSSV infection except for vp121. Of the newly identified proteins, VP466 (derived from band 16) was further characterized. The cDNA encoding VP466 was expressed in Escherichia coli as a Glutathione S-transferase (GST) fusion protein. Specific antibody was generated with the purified GST-VP466 fusion protein. Western-blot showed that the mouse anti-GST-VP466 antibody specifically bound to a 51kDa protein of WSSV. Immunogold labeling revealed that VP466 protein is a component of the viral envelope. Results in this investigation thus proved the effectiveness of proteomic approaches for discovering new proteins of WSSV.
Revised on February 5, 2002
Accepted on February 15, 2002
Proteomic analysis of shrimp white spot syndrome viral proteins and characterization of a novel envelope protein VP466
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