Assays for Oxidative Responses of Fusarium graminearum Strains to Superoxide Radicals
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Author:
Date:
2018-09-05
[Abstract] The ascomycete fungus Fusarium graminearum is a major causal agent of Fusarium head blight (FHB), a devastating disease affecting small grains cereals worldwide. To better understand the pathogenesis of this fungus, we provide here an easy-to-use protocol to examine the sensitivity of the wild-type and mutant strains of F. graminearum to oxidative stress from superoxide anions (O2•-) generated by menadione. Similarly, this assay can also be used to detect other stress responses of different fungal strains to various stress agents. The change in stress response of a mutant can offer a clue for the biological function of mutated genes.
[摘要] 子囊菌真菌 Fusarium graminearum 是 Fusarium 头枯病(FHB)的主要致病因子,FHB是一种影响全世界小粒谷物的破坏性疾病。 为了更好地了解这种真菌的发病机理,我们在此提供了一种易于使用的方案来检测 F的野生型和突变株的敏感性。 禾谷镰刀菌对由甲萘醌产生的超氧阴离子(O2 • - )的氧化应激。 类似地,该测定也可用于检测不同真菌菌株对各种应激物的其他应激反应。 突变体的应激反应的变化可以为突变基因的生物学功能提供线索。
【背景】子囊菌真菌 Fusarium graminearum (以前也被称为 Gibberella zeae 的性状)不仅是 Fusarium 头枯病和幼苗的主要致病因子小麦和大麦枯萎病,也是玉米赤霉病茎腐病的重要致病因子之一(Dal Bello et al。,2002; Bai and Shaner,2004; Kazan et al。,2012)。除了导致谷物大量减产外,这种真菌还会产生影响人类和动物健康的真菌毒素。因此,这种真菌受到广泛关注,在所有研究的植物病原真菌中排名第四(Dean et al。,2012)。
F。禾本科植物对死有机物进行过冬,特别是对受感染的小粒和玉米作物残留物进行过度研究。为了在如此广泛的环境中生存, F. graminearum ...
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Generation of Fusarium graminearum Knockout Mutants by the Split-marker Recombination Approach
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Author:
Date:
2018-08-20
[Abstract] Fusarium graminearum is a destructive phytopathogen and shows an impressive metabolic diversity. Gene deletion is an important and useful approach for gene function study. Here we present a protocol for generating gene deletion mutant by applying “split-marker” deletion strategy (Catlett et al., 2003) with PEG-mediated protoplast transformation (Yuan et al., 2008; Martín, 2015).
[摘要] 禾谷镰刀菌是一种破坏性的植物病原体,具有令人印象深刻的代谢多样性。 基因缺失是基因功能研究的重要且有用的方法。 在这里,我们提出了一个协议,通过应用“分裂标记”删除策略(Catlett et al。,2003)与PEG介导的原生质体转化(Yuan 等。,2008;Martín,2015)。
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HCV Reporter System (Viral Infection-Activated Split-Intein-Mediated Reporter System) for Testing Virus Cell-to-cell Transmission ex-vivo
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Author:
Date:
2018-08-05
[Abstract] Hepatitis C virus (HCV) spread involves two distinct entry pathways: cell-free transmission and cell-to-cell transmission. Cell-to-cell transmission is not only an efficient way for viruses to spread but also an effective method for escaping neutralizing antibodies. We adapted the viral infection-activated split-intein-mediated reporter system (VISI) and developed a straightforward model for Live-cell monitoring of HCV cell-to-cell transmission ex-vivo: co-culture of HCV infected donor cells (red signal) with uninfected recipient cells (green signal) and elimination of the cell-free transmission by adding potent neutralizing antibody AR3A in the supernatant. With this model, the efficiency of cell-to-cell transmission can be evaluated by counting the number of foci designated by ...
[摘要] 丙型肝炎病毒(HCV)传播涉及两种不同的进入途径:无细胞传播和细胞间传播。 细胞间传播不仅是病毒传播的有效方式,也是逃避中和抗体的有效方法。 我们采用了病毒感染激活的分裂 - 内含肽介导的报告系统(VISI),并开发了一种直接模型,用于活细胞监测HCV细胞间传递离体:共培养 HCV感染的供体细胞(红色信号)与未感染的受体细胞(绿色信号)和通过在上清液中加入有效的中和抗体AR3A消除无细胞的传递。 利用该模型,可以通过计数受体细胞的绿色信号指定的病灶数来评估细胞间传递的效率。
【背景】越来越多的证据证明病毒可以在受感染的组织中使用不同的传播途径(Sattentau,2008; Zhong et al。,2013)。对于HCV传播,无细胞传播和细胞间传播均可介导肝细胞之间的病毒转移。虽然无细胞传播引发HCV感染,但认为细胞 - 细胞传递直接将HCV转移至相邻的肝细胞。它提供了抵抗中和抗体并有助于病毒持久性的极好方法(Brimacombe et al。,2011; Xiao et al。,2014)。之前的文章也证明了一些促进细胞传递的宿主因子,如清道夫受体BI(SR-BI),CD81,紧密连接蛋白claudin-1(CLDN1),Occludin(OCLN),表皮生长因子受体(EGFR)。 (Witteveldt et al。,2009; ...
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