Cloning,expression and subcellular distribution of a <Emphasis Type="Italic">Rana grylio</Emphasis> virus late gene encoding ERV1 homologue |
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Authors: | Fei Ke Zhe Zhao Qiya Zhang |
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Institution: | (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 |
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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.
The sequence reported in this paper has been deposited in GenBank with the accession number, EU239358. |
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Keywords: | Rana grylio virus (RGV) Iridovirus ERV1 Late viral gene RNAi |
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