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Refrigerated incubator shaker

New BrunswickTM

Company: Eppendorf
Catalog#: Innova® 4330
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Infection of Nicotiana benthamiana Plants with Potato Virus X (PVX)
Author:
Date:
2016-12-20
[Abstract]  Potato Virus X (PVX) is the type member of Potexvirus genus, a group of plant viruses with a positive-strand RNA genome (~6.4 kb). PVX is able to establish compatible infections in Nicotiana benthamiana, a commonly used host in plant virology, leading to mild symptoms, such as chlorotic mosaic and mottling. PVX has been widely used as a viral vector for more than two decades (Chapman et al., 1992; Baulcombe et al., 1995; Aguilar et al., 2015). It provides a feasible means for the systemic expression in plants of heterologous proteins, such as avirulence factors, proteins with pharmacological properties, etc., (Hammond-Kosack et al., 1995; Gleba et al., 2014), and also as a tool to help decipher the function ... [摘要]  马铃薯病毒X(PVX)是Potexvirus属(一组具有正链RNA基因组(〜6.4kb)的植物病毒)的类型成员。 PVX能够在烟草烟草(Nicotiana benthamiana)中建立相容的感染,这是常用于植物病毒学的宿主,导致轻度症状,如褪色马赛克和斑驳。 PVX已经被广泛用作病毒载体二十多年(Chapman等人,1992; Baulcombe等人,1995; Aguilar等人。它为异源蛋白质植物中的全身表达提供了可行的手段,例如无毒性因子,具有药理学性质的蛋白质,等等(Hammond-Kosack等人, 1995; Gleba等人,2014),并且还作为通过病毒诱导的基因沉默(VACS)(Lacomme和Chapman,2008)帮助解释植物中基因功能的工具。摩擦(A)和农用过滤(B)的两种不同的方案,即允许PVX在N中的有效乘法和传播。本文详细描述本文。摩擦方法需要先前感染的液体,并且通过诱导对叶组织的机械损伤来实现感染,从而允许病毒颗粒穿透植物表面。农杆菌过滤需要先前修饰的农杆菌携带并将PVX序列递送到植物细胞中。将土壤杆菌生长直至饱和,并用注射器将其浸入植物组织中建立感染。这两种方法中的任何一种都可以成功应用,选择应主要基于材料和时间的可用性,但建议在使用嵌合病毒时使用农杆菌滤过。

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DNA Damage Sensitivity Assays in Caenorhabditis elegans
Author:
Date:
2015-06-05
[Abstract]  C. elegans has served as a genetically tractable multicellular model system to examine DNA damage-induced genotoxic stress which threatens genome integrity. Importantly, the high degree of conservation shared between worms and humans offers the advantage that findings about DNA damage-induced cell cycle arrest/checkpoint response and DNA double-strand break repair in worms are applicable to human studies. Here, we describe simple DNA damage sensitivity assays to quantify the response of C. elegans to diverse types of DNA damaging agents. These assays have provided important insights into the mechanisms of function for factors such as ZTF-8 that are involved in DNA damage repair and response in the C. elegans germline. These DNA damage sensitivity assays rely on ... [摘要]  C。 elegans 作为一种遗传易损的多细胞模型系统,以检查DNA损伤诱导的基因毒性应激,这威胁着基因组的完整性。重要的是,蠕虫和人类之间共享的高程度的保护提供了关于DNA损伤诱导的细胞周期停滞/检查点反应和DNA双链断裂修复蠕虫中的发现适用于人类研究的优势。在这里,我们描述简单的DNA损伤灵敏度测定以量化C的反应。 elegans 到各种类型的DNA损伤剂。这些测定提供了对涉及DNA损伤修复和在C中的应答的因子如ZTF-8的功能的机制的重要见解。 elegans 种系。这些DNA损伤敏感性测定依赖于卵或幼虫致死性的直接读出,并涉及使用各种DNA损伤剂。我们使用产生DNA双链断裂(DSB)的γ-照射(γ-IR),诱导单链断裂的喜树碱(CPT),氮芥(HN <2>),其产生链间交联(ICL),羟基脲(HU),其导致复制叉停止,从而防止DNA合成,和UV-C,其引起光产物(嘧啶二聚体)。参见表1.在例如突变体与野生型相比,在各种DNA损伤剂中观察到的相对灵敏度/抗性之间的比较允许关于潜在修复途径受影响的推论。

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