Development and Characterization of a cDNA-Launch Recombinant Simian Hemorrhagic Fever Virus Expressing Enhanced Green Fluorescent Protein: ORF 2b' Is Not Required for In Vitro Virus Replication.
Development and Characterization of a cDNA-Launch Recombinant Simian Hemorrhagic Fever Virus Expressing Enhanced Green Fluorescent Protein: ORF 2b' Is Not Required for In Vitro Virus Replication.
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表达增强型绿色荧光蛋白的 cDNA 启动重组猿猴出血热病毒的开发和表征：体外病毒复制不需要 ORF 2b'。
DOI：
10.3390/v13040632
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发表时间：
2021-04-07
期刊：
影响因子：
--
通讯作者：
Kuhn JH
中科院分区：
文献类型：
--
作者：
Cai Y;Yu S;Fang Y;Bollinger L;Li Y;Lauck M;Postnikova EN;Mazur S;Johnson RF;Finch CL;Radoshitzky SR;Palacios G;Friedrich TC;Goldberg TL;O'Connor DH;Jahrling PB;Kuhn JH
猴出血热病毒（SHFV）在猕猴中引起急性致命疾病。我们构建了一种单质粒cdna启动的SHFV感染克隆（rSHFV），并对其进行了修饰，以挽救表达rSHFV- egfp的增强型绿色荧光蛋白，可用于快速定量检测感染。SHFV在体外具有狭窄的细胞趋向性，只有grivet MA-104细胞系和少数其他grivet细胞系对病毒粒子进入敏感并允许感染。利用rSHFV-eGFP，我们证明了一种啮齿动物和三种猿类细胞系也完全支持SHFV复制，而55种人类细胞系、11种蝙蝠细胞系和三种啮齿动物细胞不支持SHFV复制。有趣的是，一些人类和其他哺乳动物细胞系在转染rSHFV-eGFP cdna发射质粒后，对SHFV感染具有明显抗性。为了进一步证明传染性克隆系统的研究潜力，我们在8个病毒开放阅读帧（orf）中引入了停止密码子。该方法表明，至少有一种ORF，即ORF 2b '，对于SHFV的体外复制是必不可少的。我们的原理验证实验表明，rSHFV-eGFP是阐明SHFV未被充分研究的分子生物学的有用工具。
Simian hemorrhagic fever virus (SHFV) causes acute, lethal disease in macaques. We developed a single-plasmid cDNA-launch infectious clone of SHFV (rSHFV) and modified the clone to rescue an enhanced green fluorescent protein-expressing rSHFV-eGFP that can be used for rapid and quantitative detection of infection. SHFV has a narrow cell tropism in vitro, with only the grivet MA-104 cell line and a few other grivet cell lines being susceptible to virion entry and permissive to infection. Using rSHFV-eGFP, we demonstrate that one cricetid rodent cell line and three ape cell lines also fully support SHFV replication, whereas 55 human cell lines, 11 bat cell lines, and three rodent cells do not. Interestingly, some human and other mammalian cell lines apparently resistant to SHFV infection are permissive after transfection with the rSHFV-eGFP cDNA-launch plasmid. To further demonstrate the investigative potential of the infectious clone system, we introduced stop codons into eight viral open reading frames (ORFs). This approach suggested that at least one ORF, ORF 2b’, is dispensable for SHFV in vitro replication. Our proof-of-principle experiments indicated that rSHFV-eGFP is a useful tool for illuminating the understudied molecular biology of SHFV.
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影响因子：
5
作者：
Brinton MA;Di H;Vatter HA
通讯作者：
Vatter HA
Nidoviral家族Arteriviridae的重组和扩展。
影响因子：
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作者：
Kuhn JH;Lauck M;Bailey AL;Shchetinin AM;Vishnevskaya TV;Bào Y;Ng TF;LeBreton M;Schneider BS;Gillis A;Tamoufe U;Diffo Jle D;Takuo JM;Kondov NO;Coffey LL;Wolfe ND;Delwart E;Clawson AN;Postnikova E;Bollinger L;Lackemeyer MG;Radoshitzky SR;Palacios G;Wada J;Shevtsova ZV;Jahrling PB;Lapin BA;Deriabin PG;Dunowska M;Alkhovsky SV;Rogers J;Friedrich TC;O'Connor DH;Goldberg TL
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Johnson, Reed F.
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3.7
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通讯作者：
O'Connor DH
影响因子：
5
作者：
Huang, Y. W.;Fang, Y.;Meng, X. J.
通讯作者：
Meng, X. J.