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Determination of the complete nucleotide sequences of RNA1 and RNA2 from greasy grouper (Epinephelus tauvina) nervous necrosis virus, Singapore strain

Institute of Molecular Agrobiology, The National University of Singapore, 1 Research Link, Singapore117604¹
Agri-food and Veterinary Authority, Animal Health Service Branch, Central Veterinary Laboratory, 60 Sengkang East Way, Singapore548596²
University of Tasmania, School of Biomedical Science, Locked Bag 1-320, Launceston, Tasmania 7250, Australia³
National Pingtung University of Science and Technology, Pingtung 91207, Taiwan, Republic of China⁴

Author for correspondence: Jimmy Kwang. Fax +65 8727007. e-mail kwang{at}ima.org.sg

The complete nucleotide sequences of RNA1 and RNA2 from greasy grouper (Epinephelus tauvina) nervous necrosis virus (GGNNV), Singapore strain, were determined. 5'RACE and RNA ligation were used to obtain the complete nucleotide sequences of the 5' and 3' non-coding regions (NCRs). GGNNV RNA1 was determined to be 3103 nt long, containing an ORF of 982 aa, while GGNNV RNA2 was determined to be 1433 nt long, containing an ORF of 338 aa. Both GGNNV RNAs are longer than those of other published betanodavirus sequences and the additional nucleotides were located within the NCRs. Analysis of GGNNV RNA2 revealed that it is closely related to redspotted grouper nervous necrosis virus and that both grouper viruses share the same neutralization epitope. Predicted domains for six RNA-dependent RNA polymerase motifs and two putative ORFs (proteins B1 and B2) were confirmed by sequence analysis of GGNNV RNA1.

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