| Genomic DNA Extraction and Genotyping of Dictyochloropsis Green Algae Strains
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Author:
Date:
2015-08-05
[Abstract] Dictyochloropsis is an ecologically important genus of free-living and symbiotic green algae. Representatives of this genus are horizontally transmitted among several fungi of the family Lobariaceae, thus forming photobiont-mediated guilds. This protocol is suitable for extracting DNA from algal cultures and lichen samples and for genotyping seven unlinked Dictyochloropsis reticulata microsatellite markers in a single PCR multiplex.
Figure 1. Schematic representation ...
[摘要] Dictyochloropsis是一种生态上重要的自由生活和共生绿藻类。 这个属的代表在家庭龙眼科的几种真菌之间水平传播，从而形成photobiont介导的公会。 该方案适用于从藻类培养物和地衣样品中提取DNA，以及用于在单个PCR多重中对七个未连接的微卫星标记进行基因分型。
图1.分析管道的示意图
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| Measuring Genetic Robustness in Vesicular Stomatitis Virus
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Author:
Date:
2014-03-20
[Abstract] Genetic robustness is the ability of a genome to incorporate mutations with the result of no fitness changes. Thus, more robust viruses have an increased neutral mutation rate. This property is particularly important in RNA viruses due to their high mutation rates. The most direct way of measuring robustness in vesicular stomatitis virus (VSV) is to carry out clonal analysis of populations: randomly isolating individual VSV strains (plaques), measuring the fitness of each one and generating fitness distributions (Novella et al., 2010). A second possibility is to carry out multiple ...
[摘要] 遗传稳健性是基因组掺入突变的能力，没有适应度变化的结果。因此，更强壮的病毒具有增加的中性突变率。由于它们的高突变率，这种性质在RNA病毒中特别重要。测量水泡性口炎病毒（VSV）的稳健性的最直接的方法是进行群体的克隆分析：随机分离个体VSV株（噬斑），测量每个VSV株的适合度并产生适应度分布（Novella等人， ，2010）。第二种可能性是进行重复的斑块到斑块通道的多次重复，确定后代群体中的适合度并产生健身分布（Novella等人，2010）。根据预期的差异，前者可能需要数百个测定，而后者可能需要数十次测定。第三种方法包括增加分析中的群体的突变率以扩大可能存在的任何差异，并且代替测量适合度，测量存活率（Novella等人，2013）。该方法的一个警告是生存的变化也可以通过聚合酶保真度的变化来解释。因此，重要的是进行互补实验，在这种情况下量化突变频率。
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| Fitness Determinations in Vesicular Stomatitis Virus
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Author:
Date:
2014-03-20
[Abstract] Fitness is defined as the overall replicative ability. Testing whether a mutation (or combination of mutations) has an effect on fitness often relays on determining virus production as a surrogate measurement. However, viruses do not usually replicate in a void, and evolutionary speaking, it is key to determine replicative ability compared to other viruses, e.g. the relative fitness. John Holland developed a method for vesicular stomatitis virus based on the use of a neutral genetic marker that allows to distinguish two competitors and to measure accurately the relative ratio between ...
[摘要] 健身被定义为整体的复制能力。测试突变（或突变的组合）是否对适应性有影响经常决定作为替代测量的病毒生产。然而，病毒通常不在空白中复制，并且在进化论上，确定与其他病毒相比的复制能力是关键，例如相对适合度。 John Holland开发了一种基于使用中性遗传标记物的水疱性口炎病毒的方法，所述中性遗传标记物允许区分两个竞争者并且准确地测量竞争期间两者之间的相对比率（Holland等人，1991 ）。标记是外部G糖蛋白中的突变，其对赋予对单克隆抗体I1的抗性以外的病毒没有影响。为了测量适应度，将标记的测试菌株与参考未标记的菌株混合，并使混合物感染细胞单层。通过在I1抗体存在和不存在下的噬斑测定来测量竞争前后的比率，比率的变化给出适合度值。
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