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Viral Envelope Protein 53R Gene Highly Specific Silencing and Iridovirus Resistance in Fish Cells by AmiRNA
Kim, Yu-Sin; Ke, Fei; Lei, Xiao-Ying; Zhu, Rong; Zhang, Qi-Ya; Kim, YS, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China
2010
Source PublicationPLOS ONE
ISSN1932-6203
Volume5Issue:4Pages:-
AbstractBackground: Envelope protein 53R was identified from frog Rana grylio virus (RGV), a member of the family Iridoviridae, and it plays an important role in the virus assembly. Although inhibition of iridovirus major capsid protein (MCP) by small hairpin RNAs (shRNAs) has been shown to cause resistance to viral infection in vitro, RNA interference (RNAi) to inhibit aquatic animal virus envelope protein gene product has not been reported. Methodology: We devised artificial microRNAs (amiRNAs) that target a viral envelope protein gene RGV 53R. By incorporating sequences encoding amiRNAs specific to 53R of RGV into pre-miRNA155 (pSM155) vectors, which use the backbone of natural miR-155 sequence and could intracellularly express 53R-targeted pre-amiRNAs. The pre-amiRNAs could be processed by the RNase III-like enzyme Dicer into 21-25 nt amiRNAs (amiR-53Rs) in fish cell lines. The levels of 53R expression were analyzed through real-time PCR and RGV virions assembly were observed by electronic microscopy in fish cells transfected with or without amiR-53Rs at 72 h of RGV infection. Conclusion/Significance: The results argue that viral envelope protein RGV 53R can be silenced and the virions assembly was deficient by amiR-53R-1, and further identified the first amiRNA of envelope protein gene from iridovirus that was able to cause resistance to virus infection in fish cells. The data demonstrate that the viral infection is efficiently suppressed (58%) by amiR-53R-1 targeting positon 36-57 of RGV 53R. Moreover, electron microscopic observations revealed virion assembly defect or reduced virions assembly capacity was closely correlated to expression of amiR-53R-1. Based on real time PCR of the Mx gene, we found no evidence of activation of IFN by amiR-53R-1.
KeywordRana-grylio-virus Rna Interference Paralichthys-olivaceus Functional-characterization Expression Line Inhibition System Sirna Replication
Department[Kim, Yu-Sin; Ke, Fei; Lei, Xiao-Ying; Zhu, Rong; Zhang, Qi-Ya] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China; [Kim, Yu-Sin] Kim II Sung Univ, Coll Life Sci, Pyongyang, North Korea
Subject AreaBiology ; Multidisciplinary Sciences
Funding OrganizationNational Major Basic Research Program [2009CB118704, 2010CB126303]; National Natural Science Foundation of China [30871938, 30800854]; Agriculture Commonweal Scientific Research Plan [200803013]; China Postdoctoral Science Foundation [20080440975] ; National Major Basic Research Program [2009CB118704, 2010CB126303]; National Natural Science Foundation of China [30871938, 30800854]; Agriculture Commonweal Scientific Research Plan [200803013]; China Postdoctoral Science Foundation [20080440975] ; National Major Basic Research Program [2009CB118704, 2010CB126303]; National Natural Science Foundation of China [30871938, 30800854]; Agriculture Commonweal Scientific Research Plan [200803013]; China Postdoctoral Science Foundation [20080440975] ; National Major Basic Research Program [2009CB118704, 2010CB126303]; National Natural Science Foundation of China [30871938, 30800854]; Agriculture Commonweal Scientific Research Plan [200803013]; China Postdoctoral Science Foundation [20080440975]
Indexed BySCI
Language英语
WOS IDWOS:000277079300007
Funding OrganizationNational Major Basic Research Program [2009CB118704, 2010CB126303]; National Natural Science Foundation of China [30871938, 30800854]; Agriculture Commonweal Scientific Research Plan [200803013]; China Postdoctoral Science Foundation [20080440975] ; National Major Basic Research Program [2009CB118704, 2010CB126303]; National Natural Science Foundation of China [30871938, 30800854]; Agriculture Commonweal Scientific Research Plan [200803013]; China Postdoctoral Science Foundation [20080440975] ; National Major Basic Research Program [2009CB118704, 2010CB126303]; National Natural Science Foundation of China [30871938, 30800854]; Agriculture Commonweal Scientific Research Plan [200803013]; China Postdoctoral Science Foundation [20080440975] ; National Major Basic Research Program [2009CB118704, 2010CB126303]; National Natural Science Foundation of China [30871938, 30800854]; Agriculture Commonweal Scientific Research Plan [200803013]; China Postdoctoral Science Foundation [20080440975]
Citation statistics
Cited Times:28[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/13627
Collection鱼类生物学及渔业生物技术研究中心_期刊论文
Corresponding AuthorKim, YS, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China
Recommended Citation
GB/T 7714
Kim, Yu-Sin,Ke, Fei,Lei, Xiao-Ying,et al. Viral Envelope Protein 53R Gene Highly Specific Silencing and Iridovirus Resistance in Fish Cells by AmiRNA[J]. PLOS ONE,2010,5(4):-.
APA Kim, Yu-Sin,Ke, Fei,Lei, Xiao-Ying,Zhu, Rong,Zhang, Qi-Ya,&Kim, YS, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China.(2010).Viral Envelope Protein 53R Gene Highly Specific Silencing and Iridovirus Resistance in Fish Cells by AmiRNA.PLOS ONE,5(4),-.
MLA Kim, Yu-Sin,et al."Viral Envelope Protein 53R Gene Highly Specific Silencing and Iridovirus Resistance in Fish Cells by AmiRNA".PLOS ONE 5.4(2010):-.
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