| Cycloheximide Assays to Measure Protein Degradation in vivo in Plants
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Author:
Date:
2016-09-05
[Abstract] The half-life of a protein is a characteristic property, and can be modulated by post-translational modifications, changes in subcellular localization, and/or interaction with other proteins or ligands. As one determinant of its steady-state level, a protein’s degradation represents an important distinguishing attribute relevant to its biological function. Because protein longevity cannot be elucidated from bioinformatics analyses, it must be determined empirically. Here we describe two approaches for in vivo half-life determination in plants: 1. pooled-seedling degradation assays ...
[摘要] 蛋白质的半衰期是特征性质，并且可以通过翻译后修饰，亚细胞定位的改变和/或与其他蛋白质或配体的相互作用来调节。 作为其稳态水平的一个决定因素，蛋白质的降解代表与其生物学功能相关的重要的区别属性。 由于蛋白质的寿命不能从生物信息学分析中阐明，必须根据经验确定。 这里我们描述了在植物中体内半衰期测定的两种方法：1.监测蛋白质（萤光素酶融合物或其它表位标签）的标记形式或遵循内源蛋白质的混合 - 幼苗降解测定; 2.使用荧光素酶融合蛋白的单胚芽降解测定。 这些方法的优点是它们的简单性和低成本。
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| Chemotaxis and Jumping Assays in Nematodes
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Author:
Date:
2015-09-20
[Abstract] Nematodes have sensitive olfactory perception, which is used to detect and differentiate many volatile odorants. Some odorants are attractive, others repulsive, and yet others evoke no particular response. Chemotaxis assays can be used to determine the role of certain odors in many different behaviors including foraging, predator avoidance, and mate attraction. In addition to chemotaxis, some species of nematodes in the entomopathogenic genus Steinernema can jump, which is thought to play an important role in host-seeking and dispersal (Dillman and Sternberg, 2012). Jumping and ...
[摘要] 线虫具有敏感的嗅觉感知，其用于检测和区分许多挥发性气味剂。 一些气味有吸引力，其他排斥，还有一些没有引起特别的反应。 趋化性测定可用于确定某些气味在许多不同行为中的作用，包括觅食，捕食者回避和配偶吸引。 除了趋化性之外，昆虫病原属Steinernema中的一些种类的线虫可以跳跃，这被认为在宿主寻找和分散中起重要作用（Dillman和Sternberg，2012）。 跳跃和趋化性测定已经成功地用于鉴定在多种线虫中刺激这些行为的气味剂（Bargmann等人，1993; Campbell和Kaya，1999; Hallem等人， 2011; Dillman等人，2012; Castelletto 等人，2014）。 这里基于文献的成长体提供了趋化性和跳跃测定的详细方案。
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| Quantification of HIV RNA and Human Herpesvirus DNA in Seminal Plasma
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Author:
Date:
2015-05-05
[Abstract] Multiple viruses can co-infect the genital tract, modifying the immunologic and virologic milieu and possibly playing a role in viral transmission and pathogenesis. The aim of our studies has been to understand the complex relationships between HIV-1 RNA, and multiple human herpesviruses known to frequently replicate in the genital tract of HIV-infected men (i.e. cytomegalovirus [CMV], Epstein Bar virus [EBV], herpes simplex virus [HSV] types 1 and 2, and human herpesviruses [HHV] 6, 7 and 8) (Gianella et al., 2013a; Gianella et al., 2013b; Gianella et al., ...
[摘要] 多种病毒可以共感染生殖道，修改免疫学和病毒学环境，并可能在病毒传播和发病中起作用。 我们的研究的目的是了解HIV-1 RNA和已知在HIV感染的男性生殖道中经常复制的多种人疱疹病毒之间的复杂关系（即巨细胞病毒[CMV]，Epstein 条形病毒[EBV]，单纯疱疹病毒[HSV] 1型和2型，以及人疱疹病毒[HHV] 6,7和8）（Gianella等人，2013a; Gianella等人 。，2013b; Gianella 等人，2013c; Gianella ，2014）。 该方案设计用于收集和处理男性生殖器分泌（GS），并使用定量实时PCR技术从精浆中分离并进一步定量7个HHV的HIV RNA和DNA。
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