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Cloning, expression and subcellular distribution of a Rana grylio virus late gene encoding ERV1 homologue
Ke, Fei; Zhao, Zhe; Zhang, Qiya; Zhang, QY, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China
2009-09-01
Source PublicationMOLECULAR BIOLOGY REPORTS
ISSN0301-4851
Volume36Issue:7Pages:1651-1659
AbstractAn 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.; 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.
SubtypeArticle
KeywordRana Grylio Virus (Rgv) Iridovirus Erv1 Late Viral Gene Rnai
Department[Ke, Fei; Zhao, Zhe; Zhang, Qiya] Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China
Subject AreaBiochemistry & Molecular Biology
DOI10.1007/s11033-008-9365-6
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationNational 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] ; 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] ; 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] ; 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]
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology
WOS SubjectBiochemistry & Molecular Biology
WOS IDWOS:000268496800002
WOS KeywordMULTIPLE SEQUENCE ALIGNMENT ; FISH CELL-LINE ; FAMILY IRIDOVIRIDAE ; NONSTRUCTURAL PROTEIN ; RNA INTERFERENCE ; IRIDESCENT VIRUS ; THIOREDOXIN ; INHIBITION ; MORPHOGENESIS ; TRANSCRIPTION
Funding OrganizationNational 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] ; 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] ; 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] ; 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]
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/7628
Collection期刊论文
Corresponding AuthorZhang, QY, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China
AffiliationChinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Ke, Fei,Zhao, Zhe,Zhang, Qiya,et al. Cloning, expression and subcellular distribution of a Rana grylio virus late gene encoding ERV1 homologue[J]. MOLECULAR BIOLOGY REPORTS,2009,36(7):1651-1659.
APA Ke, Fei,Zhao, Zhe,Zhang, Qiya,&Zhang, QY, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Grad Sch, Wuhan 430072, Peoples R China.(2009).Cloning, expression and subcellular distribution of a Rana grylio virus late gene encoding ERV1 homologue.MOLECULAR BIOLOGY REPORTS,36(7),1651-1659.
MLA Ke, Fei,et al."Cloning, expression and subcellular distribution of a Rana grylio virus late gene encoding ERV1 homologue".MOLECULAR BIOLOGY REPORTS 36.7(2009):1651-1659.
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