Molecular characterization of three Rana grylio virus (RGV) isolates and Paralichthys olivaceus lymphocystis disease virus (LCDV-C) in iridoviruses

Zhang, Qi-Ya; Zhao, Zhe; Xiao, Feng; Li, Zhengqiu; Gui, Jian‐Fang

doi:10.1016/j.aquaculture.2005.05.012

Cited by 34 publications

(29 citation statements)

References 29 publications

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“…Rana grylio virus (RGV) is a pathogenic agent that results in greater than 90 % mortality in cultured pig frog (Rana grylio), which was the first iridovirus isolated in China in 1995 [14,15]. RGV has been recognized as a member of the family Iridoviridae and is closely related to FV3, based on previous studies on morphogenesis, cellular interaction, antigenicity, restriction fragment length polymorphism (RFLP) and major capsid protein (MCP) sequence similarity [15][16][17][18]. To date, some genes of RGV have been identified and characterized, such as 3b-hydroxysteroid dehydrogenase (3b-HSD), deoxyuridine triphosphatase (dUTPase), an envelope protein gene (53R), thymidine kinase (TK) and a gene belonging to the ''essential for respiration and viability'' family (ERV1) [19][20][21][22][23][24].…”

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confidence: 99%

Sequencing and analysis of the complete genome of Rana grylio virus (RGV)

Zhu

et al. 2012

Arch Virol

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Infection with Rana grylio virus (RGV), an iridovirus isolated in China in 1995, resulted in a high mortality rate in frogs. The complete genome sequence of RGV was determined and analyzed. The genomic DNA was 105,791 bp long, with 106 open reading frames (ORFs). Dot plot analysis showed that the gene order of RGV shared colinearity with three completely sequenced ranaviruses. A phylogenetic tree was constructed based on concatenated sequences of iridovirus 26 core-gene-encoded proteins, and the result showed high bootstrap support for RGV being a member of the genus Ranavirus and that iridoviruses of other genera also clustered closely. A microRNA (miRNA) prediction revealed that RGV could encode 18 mature miRNAs, many of which were located near genes associated with virus replication. Thirty-three repeated sequences were found in the RGV genome. These results provide insight into the genetic nature of RGV and are useful for laboratory diagnosis for vertebrate iridoviruses.

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Sequencing and analysis of the complete genome of Rana grylio virus (RGV)

Zhu

et al. 2012

Arch Virol

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“…of one at various times post-infection (p.i.) (2, 4, 6, 8, 12, 16, 24, 36 and 48 h) or mock infected, and was subjected to Western blot analysis as described previously [31]. Anti-TK serum was used as the primary antibody at a dilution of 1:500, followed by alkaline phosphatase-conjugated goat anti-mouse IgG (H?L) antibody at a dilution of 1:1000 (Vector laboratories inc) as the secondary antibody.…”

Section: Western Blotting and Drug Inhibition Assaymentioning

confidence: 99%

“…Rana grylio virus (RGV) is a pathogen of cultured pig frog (Rana grylio) and bears significant similarity to frog virus 3 (FV3), the type species of the genus Ranavirus in the family [29][30][31]. Recently, individual proteins of RGV have been investigated to understand their roles in virus infection [32][33][34].…”

Section: Introductionmentioning

confidence: 99%

Rana grylio virus thymidine kinase gene: an early gene of iridovirus encoding for a cytoplasmic protein

et al. 2009

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The presence of thymidine kinase (TK) is a feature of many large DNA viruses. Here, a TK gene homologue was cloned and characterized from Rana grylio virus (RGV), a member of family Iridoviridae. RGV TK encodes a protein of 195 aa with a predicted molecular mass of 22.1 kDa. Homologues of the protein were present in all the currently sequenced iridoviruses, and phylogenetic analysis showed that it was much close to cellular TK type 2 (TK2), deoxycytidine kinase (dCK) and deoxyguanosine kinase (dGK). Subsequently, Western blotting revealed TK expression increased with time from 6 h postinfection in RGV-infected cells. Using drug inhibition analysis by protein synthesis inhibitor (cycloheximide) and DNA replication inhibitor (cytosine arabinofuranoside), RGV TK was classified as the early expression gene during in vitro infection. Subcellular localization by TK-GFP fusion protein expression and immunofluorescence staining showed RGV TK was an exclusively cytoplasmic protein in fish cells. Collectively, current data indicate that RGV TK was an early gene of iridovirus which encoded a cytoplasmic protein in fish cells.

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“…The procedures of virus propagation and purification were according to previous reports (Zhang et al, 2006;Tao et al, 2007). The virus pellet was resuspended in TE buffer (10 mM Tris-Cl, 1 mM EDTA, pH 7.4) and stored at −20 • C until use.…”

Section: Virus Propagation Purification and Rna Preparationmentioning

confidence: 99%