| Measuring Cyanobacterial Metabolism in Biofilms with NanoSIMS Isotope Imaging and Scanning Electron Microscopy (SEM)
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Author:
Date:
2017-05-05
[Abstract] To advance the understanding of microbial interactions, it is becoming increasingly important to resolve the individual metabolic contributions of microorganisms in complex communities. Organisms from biofilms can be especially difficult to separate, image and analyze, and methods to address these limitations are needed. High resolution imaging secondary ion mass spectrometry (NanoSIMS) generates single cell isotopic composition measurements, and can be used to quantify incorporation and exchange of an isotopically labeled substrate among individual organisms. Here, incorporation of cyanobacterial extracellular organic matter (EOM) by members of a cyanobacterial mixed species biofilm is used as a model to illustrate this method. Incorporation of stable isotope labeled (15N and 13 ...
[摘要] 为了提高对微生物相互作用的理解，解决复杂群落中微生物代谢贡献的重要性越来越重要。生物膜的生物体特别难以分离，形成和分析，需要解决这些局限性的方法。高分辨率成像二次离子质谱（NanoSIMS）产生单细胞同位素组成测量，可用于量化在各生物体内同位素标记的底物的掺入和交换。在这里，使用由蓝藻混合物种生物膜的成员掺入的蓝细菌胞外有机物（EOM）作为模型来说明该方法。通过两组（蓝细菌和相关的异养微生物）掺入稳定同位素（ 15 N和 13 C）EOM的量化。描述了用于定量生物膜中稳定同位素标记的EOM摄取的样品的生成，制备和分析方法。
背景 与NanoSIMS（“NanoSIP”）结合的稳定同位素标记是一种确定的稳定同位素标记底物掺入到单个微生物细胞中的方法，然后可将其推断以估计细胞群体的掺入（例如，Lechene et al。 ，2006和Woebken等人，2012）。可以将多个稳定同位素标记（例如， 13 C和 15 ...
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| Preparation of Bacillus subtilis Cell Lysates and Membranes
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Author:
Date:
2014-01-20
[Abstract] A common feature of every eukaryotic and prokaryotic cell is that they exhibit a plasma membrane. In Bacillus subtilis (B. subtilis) roughly 25% of all proteins are putative trans- or membrane associated proteins. Here we describe a relatively simple method to separate and prepare membrane and cytosolic proteins by ultra-centrifugation.
[摘要] 每个真核和原核细胞的共同特征是它们表现出质膜。 在枯草芽孢杆菌（<枯草芽孢杆菌）中，大约25％的蛋白质是推定的跨膜或膜相关蛋白。 在这里，我们描述了一种相对简单的方法，通过超离心分离和制备膜和胞质蛋白
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| Isolation of the Secretome from Bacillus subtilis
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Author:
Date:
2014-01-20
[Abstract] Bacteria are commonly known to secret proteins in large amounts into the surrounding environment in high concentrations via various pathways. These proteins can be involved in numerous processes like cell-cell communication, exopolymer formation but also metabolic active enzymes are secreted that are interesting for industrial production of proteins. One of the most regularly used organisms for industrial protein production is the Gram-positive bacterium Bacillus subtilis (B. subtilis). Here we describe a protocol that can be used to quantitatively and qualitatively analyze secreted proteins from B. subtilis.
[摘要] 通常已知细菌通过各种途径以高浓度将蛋白质大量地分泌到周围环境中。 这些蛋白质可以参与许多过程，例如细胞 - 细胞通讯，外聚体形成，但是也分泌代谢活性酶，其对于蛋白质的工业生产是有趣的。 用于工业蛋白质生产的最经常使用的生物体之一是革兰氏阳性细菌枯草芽孢杆菌（<枯草芽孢杆菌）。 在这里我们描述可以用于定量和定性分析从分泌蛋白质的协议。="">
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