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Vero cells

Vero

Company: ATCC
Catalog#: CCL-81
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Random Insertional Mutagenesis of a Serotype 2 Dengue Virus Clone
Author:
Date:
2018-08-20
[Abstract]  Protein tagging is a powerful method of investigating protein function. However, modifying positive-strand RNA virus proteins in the context of viral infection can be particularly difficult as their compact genomes and multifunctional proteins mean even small changes can inactivate or attenuate the virus. Although targeted approaches to functionally tag viral proteins have been successful, these approaches are time consuming and inefficient. A strategy that has been successfully applied to several RNA viruses is whole-genome transposon insertional mutagenesis. A library of viral genomes, each containing a single randomly placed small insertion, is selected by passaging in cell culture and the insertion sites can be identified using Next Generation Sequencing (NGS). Here we describe a ... [摘要]  蛋白质标记是研究蛋白质功能的有效方法。然而,在病毒感染的情况下修饰正链RNA病毒蛋白可能特别困难,因为它们的紧密基因组和多功能蛋白意味着即使很小的变化也可以使病毒失活或减弱。尽管功能性标记病毒蛋白的靶向方法已经成功,但这些方法耗时且效率低。已经成功应用于几种RNA病毒的策略是全基因组转座子插入诱变。通过细胞培养中的传代选择病毒基因组文库,每个文库含有单个随机放置的小插入,并且可以使用下一代测序(NGS)鉴定插入位点。在这里,我们描述了用于登革病毒16681株血清型2的转座子诱变的方案。含有短随机放置插入物的突变登革病毒文库通过哺乳动物细胞传代,插入由有活力后代的NGS定位。该方案分为四个阶段:登革热cDNA克隆的转座子诱变,病毒基因组转染到允许细胞,分离病毒后代基因组和测序文库制备。

【背景】 ...

Virucidal and Neutralizing Activity Tests for Antiviral Substances and Antibodies
Author:
Date:
2018-05-20
[Abstract]  In a narrow definition, virucidal activity represents the activity by which to interact with and physically disrupt viral particles. In a broad definition, it includes the activity by which to functionally inhibit (neutralize) viral infectivity without apparent morphological alterations of the viral particles. The viral infectivity can be measured in cell culture system by means of plaque assay, infectious focus assay, 50% tissue culture infectious dose (TCID50) assay, etc. Morphologically, disruption of viral particles can be demonstrated by negative staining electron microscopic analysis of viral particles. In this article, we describe methods to assess virucidal activity in a broad definition. [摘要]  在狭义定义中,杀病毒活性表示与病毒颗粒相互作用和物理破坏的活性。 在一个广义的定义中,它包括了功能上抑制(中和)病毒感染性而没有病毒颗粒的明显形态改变的活性。 可以通过噬菌斑测定,感染性聚焦测定,50%组织培养感染剂量(TCID 50)测定,等等在细胞培养系统中测量病毒感染性。 形态学上,病毒颗粒的破坏可以通过病毒颗粒的负染色电子显微镜分析来证明。 在这篇文章中,我们描述了用广义定义评估杀病毒活性的方法。

【背景】病毒是细胞内寄生虫,它们劫持宿主细胞机制来复制它们自己的基因组。在病毒生命周期的最初阶段,感染性病毒颗粒附着(结合)到靶细胞表面上的称为病毒受体的特定宿主蛋白,然后病毒渗透(内化和/或融合)到细胞的细胞内区室宿主细胞,其中病毒生命周期的后续步骤继续产生后代病毒体(Scheel和Rice,2013)。

狭义定义中的杀病毒活性表示与病毒颗粒相互作用和物理破坏的活性。在一个广义的定义中,它包括与病毒感染性相互作用并在功能上抑制(中和)病毒感染性而没有病毒颗粒的明显形态改变的活性,如抗体介导的中和的情况。

最近我们报道了从蛇毒(Chen等,2017)获得的分泌型磷脂酶Aβ的异构体和来自蝎毒(El-Bitar ...

Lentiviral Knockdown of Transcription Factor STAT1 in Peromyscus leucopus to Assess Its Role in the Restriction of Tick-borne Flaviviruses
Author:
Date:
2017-12-05
[Abstract]  Cellular infection with tick-borne flaviviruses (TBFVs) results in activation of the interferon (IFN) signaling pathway and subsequent upregulation of numerous genes termed IFN stimulated genes (ISGs) (Schoggins et al., 2011). Many ISGs function to prevent virus pathogenesis by acting in a broad or specific manner through protein-protein interactions (Duggal and Emerman, 2012). The potency of the IFN signaling response determines the outcome of TBFV infection (Best, 2016; Carletti et al., 2017). Interestingly, data from our lab show that TBFV replication is significantly restricted in cells of the reservoir species Peromyscus leucopus thereby suggesting a potent antiviral response (Izuogu et al., 2017). We assessed the relative contribution of IFN ... [摘要]  蜱传黄热病病毒(TBFV)的细胞感染导致干扰素(IFN)信号传导途径的激活和随后称为IFN刺激基因(ISG)(Schoggins等人,2011)的众多基因的上调。许多ISG通过蛋白质 - 蛋白质相互作用以广泛或特定的方式起作用来防止病毒发病(Duggal和Emerman,2012)。 IFN信号反应的效力决定了TBFV感染的结果(Best,2016; Carletti等人,2017)。有趣的是,我们实验室的数据显示TBFV复制在储库物种Peromyscus leucopus的细胞中显着受到限制,从而表明有效的抗病毒应答(Izuogu等人,2017)。我们评估干扰素信号对抗性的相对贡献。通过敲低IFN反应途径中的主要转录因子来抑制白血病。信号转导和转录激活因子1(STAT1)是专门针对在P。 leucopus细胞通过shRNA技术。我们进一步测试了基因敲低对细胞对IFN反应和限制病毒复制的能力的影响;结果表明当STAT1表达被改变时,leucopus细胞对IFN刺激的反应降低,并且对TBFV复制显着更敏感。

【背景】IFN信号是抵抗侵入宿主细胞的黄病毒的第一道防线(Robertson等人,2009; Lazear和Diamond,2015)。通过模式识别受体(PRR)检测与病毒颗粒相关的分子标记,然后通过转录因子引发下游信号从细胞释放1型IFN(Kawai ...

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