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Non-virulence of a recombinant shrimp nidovirus is associated with its non structural gene sequence and not a large structural gene deletion
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Document Title
Non-virulence of a recombinant shrimp nidovirus is associated with its non structural gene sequence and not a large structural gene deletion
Author
Gangnonngiw W., Anantasomboon G., Sang-oum W., Sriurairatana S., Sritunyalucksana K., Flegel T.W.
Name from Authors Collection
Scopus Author ID
6503848431
Affiliations
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Thailand Science Park, Klong Luang Pratumthani 12120, Thailand; Centex Shrimp, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok, 10400, Thailand; Department of Anatomy, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok, 10400, Thailand; Anatomy Unit, Department of Medical Science, Faculty of Science, Patumthani, 12000, Thailand; Department of Biotechnology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok, 10400, Thailand
Type
Article
Source Title
Virology
ISSN
00426822
Year
2009
Volume
385
Issue
1
Page
161-168
Open Access
Bronze
DOI
10.1016/j.virol.2008.10.044
Abstract
RT-PCR using a commercial kit for yellow head virus (YHV) detection in growth-retarded shrimp yielded an unusual 777 bp amplicon instead of expected amplicons of 277 bp for YHV type-1 (YHV-1) or 406 bp for YHV type-2 (YHV-2). Cloning and sequencing (GenBank EU170438) revealed approximately 80% identity to non-structural (NS) ORF1b sequences of both YHV-1 (GenBank AA083987) and YHV-2 (GenBank AF227196), indicating an atypical YHV type (A-YHV) phylogenetically equidistant from both types. An RT-PCR test specifically designed for A-YHV revealed that it was uncommon and that its occurrence in shrimp culture ponds did not correlate with growth retardation or mortality. By immunohistochemistry with YHV-specific monoclonal antibodies, the A-YHV gave positive reactions for envelope protein gp64 and capsid protein p20, but not for envelope protein gp116, even though gp116 and gp64 originate from a polyprotein of ORF3. Lack of gp116 immunoreactivity correlated with a large ORF3 deletion (GenBank EU123854) in the region of the protein targeted by an MAb against gp116. Transmission electron microscopy of A-YHV-infected shrimp revealed only unenveloped pre-virions. During manuscript revision, information received revealed that typing of YHV isolates based on sequences of ORF1b and ORF3 had yielded several geographical types, including one virulent type (YHV-1b) with an ORF3 deletion sequence that matched the sequence of A-YHV. Using these sequences and an additional A-YHV sequence (EU853170) from the ORF1b typing region, A-YHV potentially represents a recombinant between type 1b and type 5. SDS-PAGE and Western blot analysis revealed that type 1b produced a gp116 deletion protein that did not bind with the MAb or polyclonal Ab to normal gp116. Overall, the information suggested that lack of A-YHV virulence was associated with the NS gene sequence linked to ORF1b rather than the deletion in ORF3. © 2008 Elsevier Inc. All rights reserved.
Keyword
Deletion | Gp116 | Monodon slow growth syndrome | MSGS | Penaeus monodon | Recombinant | Virulence | Yellow head virus | YHV
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
License
CC BY or CC BY-NC-ND
Rights
Elsevier B.V.
Publication Source
Scopus