Envelope Protein Gene VP466-a Target for PCR Detection of White Spot Syndrome Virus in Shrimp

White spot syndrome virus (WSSV) is an enveloped and double-stranded DNA virus that belongs to the family Nimaviridae and genus Whispovirus, causing white spot disease (WSD) in shrimp. The virus is highly virulent and leads to 100% mortality within 10 days. Detection of WSSV and segregation of infected brood shrimp, post-larvae and cultured shrimp are currently considered as containment strategies to reduce the spread of WSD. This investigation describes a polymerase chain reaction method to detect WSSV in WSD infected cultured shrimp targeting VP466 gene encoding the large structural protein in virus particle. In silico homology analysis of the primer pair designed in this work clearly identified WSSV VP466 gene sequence with 100% specificity. A total of 16 shrimp samples from 16 farms were selected, where 6 shrimp samples were with characteristics WSD spot and 10 shrimp samples were asymptomatic. Among the 16 shrimp samples, 12 showed PCR positive amplifications for major envelope protein gene VP466. Sequencing of the amplicons followed by homology searching using BLAST further confirmed the presence of WSSV. Phylogenetic analysis of VP466 gene sequences showed its close proximity to the WSSV strain of Indian origin. The present study demonstrates that the envelope protein VP466 gene as a specific target for PCR detection and characterization of WSSV in WSD infected and carrier shrimpsBangladesh J Microbiol, Volume 31, Number 1-2,June-Dec 2014, pp 65-68

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Characterization of a novel envelope protein WSV010 of shrimp white spot syndrome virus and its interaction with a major viral structural protein VP24

Virology ◽

10.1016/j.virol.2007.02.030 ◽

2007 ◽

Vol 364 (1) ◽

pp. 208-213 ◽

Cited By ~ 22

Author(s):

Jing Chen ◽

Zhengjun Li ◽

Choy-Leong Hew

Keyword(s):

Envelope Protein ◽

White Spot Syndrome Virus ◽

White Spot ◽

Structural Protein ◽

Viral Structural Protein ◽

White Spot Syndrome

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Transcription and identification of an envelope protein gene (p22) from shrimp white spot syndrome virus

Journal of General Virology ◽

10.1099/0022-1317-83-2-471 ◽

2002 ◽

Vol 83 (2) ◽

pp. 471-477 ◽

Cited By ~ 55

Author(s):

Xiaobo Zhang ◽

Canhua Huang ◽

Xun Xu ◽

Choy L. Hew

Keyword(s):

Fusion Protein ◽

Envelope Protein ◽

White Spot Syndrome Virus ◽

Protein Gene ◽

Temporal Analysis ◽

White Spot ◽

Outer Envelope ◽

Reading Frame ◽

Transmission Electron ◽

White Spot Syndrome

White spot syndrome virus (WSSV) is one of the most virulent pathogens causing high mortality in shrimp. In the present study, an open reading frame (termed the p22 gene) was revealed from a WSSV cDNA library. The gene was expressed as a fusion protein with glutathione S-transferase (GST) in Escherichia coli and purified. Specific antibody was raised using the purified fusion protein (GST–P22). Temporal analysis showed that the p22 gene was a late gene. After binding between purified WSSV virions and anti-GST–P22 IgG followed by labelling with gold-labelled secondary antibody, the gold particles, under a transmission electron microscope, could be found along the outer envelope of WSSV virions. This experiment suggests that the p22 gene encodes an envelope protein of the virus.

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A novel envelope protein involved in White spot syndrome virus infection

Journal of General Virology ◽

10.1099/vir.0.80923-0 ◽

2005 ◽

Vol 86 (5) ◽

pp. 1357-1361 ◽

Cited By ~ 28

Author(s):

Ru Huang ◽

Yunli Xie ◽

Jianhong Zhang ◽

Zhengli Shi

Keyword(s):

Escherichia Coli ◽

Virus Infection ◽

Envelope Protein ◽

White Spot Syndrome Virus ◽

Open Reading Frame ◽

White Spot ◽

Structural Protein ◽

Reading Frame ◽

Intact Virion ◽

White Spot Syndrome

One open reading frame (designated vp76) from the White spot syndrome virus (WSSV) genome has the motif of a cytokine I receptor and has been identified as a structural protein. In this paper, vp76 was expressed in Escherichia coli and used to prepare a specific antibody to determine the location of the corresponding protein in the intact virion, the nucleocapsids and the envelope of WSSV. Western blotting with the VP76 antiserum confirmed that VP76 was an envelope protein of WSSV. To investigate the function of the VP76, WSSV was neutralized with the VP76-specific antiserum at different concentrations and injected intramuscularly into crayfish. The mortality curves showed that the VP76 antiserum could partially attenuate infection with WSSV, suggesting that VP76 is an envelope protein involved in WSSV infection.

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Identification and localization of a prawn white spot syndrome virus gene that encodes an envelope protein

Journal of General Virology ◽

10.1099/0022-1317-83-5-1069 ◽

2002 ◽

Vol 83 (5) ◽

pp. 1069-1074 ◽

Cited By ~ 45

Author(s):

Xiaobo Zhang ◽

Canhua Huang ◽

Xun Xu ◽

Choy L. Hew

Keyword(s):

Envelope Protein ◽

White Spot Syndrome Virus ◽

Penaeus Monodon ◽

White Spot ◽

Cdna Libraries ◽

Gene Transcript ◽

Virus Envelope ◽

Gene Encoding ◽

A Genome ◽

White Spot Syndrome

Among the important challenges to shrimp aquaculture worldwide are the diseases caused by viruses, in particular by white spot syndrome virus (WSSV), which has a genome estimated to contain 305 kb. By analysis and comparison of the WSSV genomic DNA and cDNA libraries, an ORF (vp28 gene) was identified. The gene, encoding a novel 204-amino-acid protein, was expressed in Escherichia coli and purified. A specific antibody was raised using the purified VP28 protein. After inoculation of healthy adult Penaeus monodon shrimp with WSSV, the gene transcript and VP28 protein were first detected at low levels at 6 and 18 h post-infection, respectively. These experiments suggest that it might be a late gene. Immuno-electron microscopy with gold-labelled antibody revealed that the gold particles were distributed in the outer envelope of WSSV virions and showed that vp28 encodes a virus envelope protein.

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White spot syndrome virus VP24 interacts with VP28 and is involved in virus infection

Journal of General Virology ◽

10.1099/vir.0.81570-0 ◽

2006 ◽

Vol 87 (7) ◽

pp. 1903-1908 ◽

Cited By ~ 47

Author(s):

Xixian Xie ◽

Feng Yang

Keyword(s):

Virus Infection ◽

Envelope Protein ◽

White Spot Syndrome Virus ◽

Early Stage ◽

White Spot ◽

Structural Protein ◽

Virion Envelope ◽

White Spot Syndrome ◽

Triton X

White spot syndrome virus (WSSV) is one of the most virulent pathogens causing high mortality in shrimp. Herein, the characterization of VP24, a major structural protein of WSSV, is described. When purified virions were subjected to Nonidet P-40 treatment to separate the envelopes from the nucleocapsids, VP24 was found to be present exclusively in the envelope fraction. Triton X-114 extraction also indicated that VP24 behaves as an envelope protein. Immunoelectron microscopy further confirmed that VP24 is located in the virion envelope. Far-Western experiments showed that VP24 interacts with VP28, another major envelope protein of the WSSV virion. To investigate the function of VP24, WSSV was neutralized with various amounts of anti-VP24 IgG and injected into crayfish. The results showed that anti-VP24 IgG could partially attenuate infection with WSSV. It is concluded that VP24 is an envelope protein and functions at an early stage in virus infection.

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Molecular identification of white spot syndrome virus (WSSV) and associated risk factors for white spot disease (WSD) prevalence in shrimp (Penaeus monodon) aquaculture in Bangladesh

Journal of Invertebrate Pathology ◽

10.1016/j.jip.2021.107535 ◽

2021 ◽

Vol 179 ◽

pp. 107535

Author(s):

Abu Salman Talukder ◽

Nusrat Jahan Punom ◽

Md. Mostavi Enan Eshik ◽

Mst. Khadiza Begum ◽

H.M. Rakibul Islam ◽

...

Keyword(s):

Risk Factors ◽

Molecular Identification ◽

White Spot Syndrome Virus ◽

Penaeus Monodon ◽

White Spot ◽

White Spot Disease ◽

Spot Disease ◽

White Spot Syndrome ◽

Associated Risk Factors

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Crystal structure of the C-terminal domain of envelope protein VP37 from white spot syndrome virus reveals sulphate binding sites responsible for heparin binding

Journal of General Virology ◽

10.1099/jgv.0.001611 ◽

2021 ◽

Vol 102 (6) ◽

Author(s):

Wasusit Somsoros ◽

Takeshi Sangawa ◽

Katsuki Takebe ◽

Jakrada Attarataya ◽

Kanokpan Wongprasert ◽

...

Keyword(s):

Crystal Structure ◽

Binding Sites ◽

Envelope Protein ◽

Molecular Mechanisms ◽

White Spot Syndrome Virus ◽

Significant Loss ◽

White Spot ◽

Heparin Binding ◽

Terminal Domain ◽

White Spot Syndrome

White spot syndrome virus (WSSV) is the most virulent pathogen causing high mortality and economic loss in shrimp aquaculture and various crustaceans. Therefore, the understanding of molecular mechanisms of WSSV infection is important to develop effective therapeutics to control the spread of this viral disease. In a previous study, we found that VP37 could bind with shrimp haemocytes through the interaction between its C-terminal domain and heparin-like molecules on the shrimp cells, and this interaction can also be inhibited by sulphated galactan. In this study, we present the crystal structure of C-terminal domain of VP37 from WSSV at a resolution of 2.51 Å. The crystal structure contains an eight-stranded β-barrel fold with an antiparallel arrangement and reveals a trimeric assembly. Moreover, there are two sulphate binding sites found in the position corresponding to R213 and K257. In order to determine whether these sulphate binding sites are involved in binding of VP37 to heparin, mutagenesis was performed to replace these residues with alanine (R213A and K257A), and the Surface Plasmon Resonance (SPR) system was used to study the interaction of each mutated VP37 with heparin. The results showed that mutants R213A and K257A exhibited a significant loss in heparin binding activity. These findings indicated that the sites of R213 and K257 on the C-terminal domain of envelope protein VP37 are essential for binding to sulphate molecules of heparin. This study provides further insight into the structure of C-terminal domain of VP37 and it is anticipated that the structure of VP37 might be used as a guideline for development of antivirus agent targeting on the VP37 protein.

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