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Cloning, expression and subcellular distribution of a Rana grylio virus late gene encoding ERV1 homologue

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Abstract

An essential for respiration and viability (ERV1) homologue, 88R, was cloned and characterized from Rana grylio virus (RGV). Database searches found its homologues in all sequenced iridoviruses, and sequence alignment revealed a highly conserved motif shared by all ERV1 family proteins: Cys-X-X-Cys. RT-PCR and western blot analysis revealed that 88R begins to transcribe and translate at 6 h postinfection (p.i.) and remains detectable at 48 h p.i. during RGV infection course. Furthermore, using drug inhibition analysis by a de novo protein synthesis inhibitor and a viral DNA replication inhibitor, RGV 88R was classified as a late (L) viral gene during the in vitro infection. 88R-EGFP fusion protein was observed in both the cytoplasm and nucleus of pEGFP-N3-88R transfected EPC cells. Although result of immunofluorescence is similar, 88R protein was not detected in viromatrix. Moreover, function of RGV 88R on virus replication were evaluated by RNAi assay. Nevertheless, effect of knockdown of RGV 88R expression on virus replication was not detected in cultured fish cell lines. Collectively, current data indicate that RGV 88R was a late gene of iridovirus encoding protein that distributed both the cytoplasm and nucleus.

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Acknowledgments

This work was supported by grants from the National Major Basic Research Program (2004CB117403), the National 863 High Technology Research Foundation of China (2006AA09Z445, 2006AA100309 and 20060110A4013), the National Natural Science Foundation of China (30671616 and U0631008) and the Key Technology R & D Program of China (2006BAD03B05).

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Authors and Affiliations

  1. State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Wuhan, 430072, China

    Fei Ke, Zhe Zhao & Qiya Zhang

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  1. Fei Ke
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  2. Zhe Zhao
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  3. Qiya Zhang
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Correspondence to Qiya Zhang.

Additional information

The sequence reported in this paper has been deposited in GenBank with the accession number, EU239358.

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Ke, F., Zhao, Z. & Zhang, Q. Cloning, expression and subcellular distribution of a Rana grylio virus late gene encoding ERV1 homologue. Mol Biol Rep 36, 1651–1659 (2009). https://doi.org/10.1007/s11033-008-9365-6

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