| 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 ...
[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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| An Evaluation of Cellulose Degradation Affected by Dutch Elm Disease
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Author:
Date:
2015-07-20
[Abstract] The pathogenic fungus Ophiostoma novo-ulmi spreads within the secondary xylem vessels of infected elm trees, causing the formation of vessel plugs due to tyloses and gels, which ultimately result in Dutch elm disease. Foliage discoloration, wilting and falling from the tree are typical external leaf symptoms of the disease followed by the subsequent death of sensitive trees. Cellulolytic enzymes produced by the fungus are responsible for the degradation of medium molecular weight macromolecules of cellulose, resulting in the occurrence of secondary cell wall ruptures and cracks in ...
[摘要] 病原真菌新孢子虫在受感染的榆树的次生木质部导管内扩散，导致由于菌丝和凝胶而形成血管栓塞，最终导致荷兰榆树病。叶子变色，枯萎和从树上掉下是典型的外部叶症状的疾病，随后是敏感树的死亡。由真菌产生的纤维素分解酶负责纤维素的中等分子量大分子的降解，导致在血管中发生继发性细胞壁破裂和裂纹，但是在纤维中很少发生破裂（Jurkovic等人， ，2014）。该程序的目的是通过高侵蚀性的O应变评价纤维素降解的程度。 novo-ulmi ssp。 美国× novo-ulmi 。尺寸排阻色谱法（SEC）比较了感染的和未感染的榆树之间的纤维素的分子量分布，并揭示了纤维素的大分子性状的变化，包括分子量，聚合度和多分散指数。 13 C魔幻角旋转核磁共振（ 13 C MAS NMR）光谱有助于鉴定和定量由于降解而导致的结晶和非结晶纤维素区域的损失。本文所述的方法也可以容易地用于感染各种纤维素降解真菌的其它木本植物。
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| Analysis of Sugar Component of a Hot Water Extract from Arabidopsis thaliana Pollen Tubes Using GC-EI-MS
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Author:
Date:
2015-06-20
[Abstract] Extraction with hot water is the oldest and simplest method used to recover pectin from an alcohol insoluble residue extract, although this method has not been widely used for the cell wall analysis of pollen tube, a model used to study cell wall. This protocol described this method applied for pectin extraction from 6 h-old Arabidopsis pollen tubes followed by a sugar composition analysis by gas chromatography mass spectrometry.
[摘要] 用热水提取是用于从醇不溶性残余物提取物中回收果胶的最古老和最简单的方法，尽管该方法尚未广泛用于花粉管的细胞壁分析，所述花粉管是用于研究细胞壁的模型。 该方案描述了该方法应用于从6h-旧拟南芥花粉管中提取果胶，然后通过气相色谱质谱法进行糖组成分析。
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