| Single-cell Visualization of Chromosome Transcriptional Territories by RNA-paint
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Author:
Date:
2016-09-05
[Abstract] We developed a FISH-based method to directly assess chromosome-wide transcriptional activity, thereby enabling the visualization of the actively transcribed fraction of a chromosome at the single-cell level. We applied this method to probe the activity of X-chromosomes and its instability in the context of human embryonic stem cells and cancer cells.
[摘要] 我们开发了基于FISH的方法，以直接评估染色体宽转录活性，从而使单细胞水平的染色体的积极转录部分的可视化。 我们应用这种方法探测X染色体的活动及其在人类胚胎干细胞和癌细胞的上下文中的不稳定性。
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| Assay of Arabinofuranosidase Activity in Maize Roots
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Author:
Date:
2016-03-20
[Abstract] Root is a perfect model for studying the mechanisms of plant cell growth. Along the root length, several zones where cells are at different stages of development can be visualized (Figure 1). The dissection of the root on these zones allows the investigation of biochemical and genetic aspects of different growth steps. Maize primary root is much more massive than the root of other Monocots and thus more convenient for such type of research. Plant cell wall, mainly consisting of polysaccharides, plays an important role in plant life. Therefore, measurement of plant carbohydrate content and glycoside-modifying enzyme activity in plant cells has become an important aspect in plant physiology. One of the well-documented changes of hemicelluloses molecules during elongation growth of monocots ...
[摘要] 根是研究植物细胞生长机制的完美模型。沿着根长度，可以可视化细胞处于不同发育阶段的几个区（图1）。在这些区域上的根的解剖允许调查不同生长步骤的生化和遗传方面。玉米主根比其他单根的根更大，因此这种类型的研究更方便。植物细胞壁，主要由多糖组成，在植物生活中发挥重要作用。因此，植物碳水化合物含量和糖苷修饰酶活性在植物细胞中的测量已经成为植物生理学中的重要方面。在单子叶植物细胞的延长生长期间半纤维素分子的充分证明的变化之一是葡糖醛酸阿拉伯木聚糖的阿拉伯糖取代的减少。这可能是由于该多糖的合成的变化或阿拉伯呋喃糖苷酶的作用引起的。在这里，我们描述的分光光度测量阿拉伯呋喃糖苷酶活性在玉米根中的水解发色底物（4-硝基苯基α-L-阿拉伯呋喃糖苷）的速率的协议。
图1. 四日龄黑暗生长的玉米幼苗面板）。根据Kozlova等人（2012）（右图），初级玉米根的不同区和细胞发育的相应阶段。
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| Purification of Bacterial RNA from Infected Macrophages
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Author:
Date:
2015-11-20
[Abstract] Studying the transcriptome of bacterial pathogens during infection is a very informative and effective tool for discovering genes that contribute to successful infection. However, isolating bacterial RNA from infected cells or tissues is a challenging process due to the much higher amounts of host RNA in the lysates of infected cells. We have optimized a method for isolating RNA of Listeria monocytogenes (L. monocytogenes) bacteria infecting bone marrow derived macrophage cells (BMDM). After infection, we lyse the cells and filter the lysates through 0.45 µm filters to discard most of the host proteins and RNA. Next, we resuspend the bacteria and extract RNA following DNase treatment. The extracted RNA is suitable for gene expression analysis by real-time PCR or ...
[摘要] 在感染期间研究细菌病原体的转录组是一个非常有益的和有效的工具，用于发现有助于成功感染的基因。然而，从感染的细胞或组织中分离细菌RNA是一个挑战性的过程，因为感染细胞裂解物中宿主RNA的量高得多。我们已经优化了用于分离感染骨髓来源的巨噬细胞（BMDM）的单核细胞增生性李斯特菌（（单核细胞增生李斯特氏菌）细菌）的RNA的方法。感染后，我们裂解细胞并通过0.45μm过滤器过滤裂解物以丢弃大多数宿主蛋白和RNA。接下来，我们重新悬浮细菌，并在DNase处理后提取RNA。提取的RNA适合于通过实时PCR或微阵列的基因表达分析。我们已经在我们对感染期间的单核细胞增生李斯特氏菌基因调节的体外研究中成功地采用了该方案（Lobel等人，2015; Lobel >，2012; Kaplan Zeevi等人，2013; Rabinovich等人，2012）。
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