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学科主题: Biochemistry & Molecular Biology
题名: Cloning, expression and subcellular distribution of a Rana grylio virus late gene encoding ERV1 homologue
作者: Ke, Fei1; Zhao, Zhe1; Zhang, Qiya1
通讯作者: Zhang, QY, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China
关键词: Rana grylio virus (RGV) ; Iridovirus ; ERV1 ; Late viral gene ; RNAi
刊名: MOLECULAR BIOLOGY REPORTS
发表日期: 2009-09-01
DOI: 10.1007/s11033-008-9365-6
卷: 36, 期:7, 页:1651-1659
收录类别: SCI
文章类型: Article
部门归属: [Ke, Fei; Zhao, Zhe; Zhang, Qiya] Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
资助者: National Major Basic Research Program [2004CB117403]; National 863 High Technology Research Foundation of China [2006AA09Z445, 2006AA100309, 20060110A4013]; National Natural Science Foundation of China [30671616, U0631008]; Key Technology R & D Program of China [2006BAD03B05]
类目[WOS]: Biochemistry & Molecular Biology
研究领域[WOS]: Biochemistry & Molecular Biology
摘要: 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.
英文摘要: 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.
关键词[WOS]: MULTIPLE SEQUENCE ALIGNMENT ; FISH CELL-LINE ; FAMILY IRIDOVIRIDAE ; NONSTRUCTURAL PROTEIN ; RNA INTERFERENCE ; IRIDESCENT VIRUS ; THIOREDOXIN ; INHIBITION ; MORPHOGENESIS ; TRANSCRIPTION
语种: 英语
WOS记录号: WOS:000268496800002
ISSN号: 0301-4851
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/7628
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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作者单位: 1.Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China

Recommended Citation:
Ke, Fei; Zhao, Zhe; Zhang, Qiya.Cloning, expression and subcellular distribution of a Rana grylio virus late gene encoding ERV1 homologue,MOLECULAR BIOLOGY REPORTS,2009,36(7):1651-1659
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