| Extraction of Small Molecules from Fecal Samples and Testing of Their Activity on Microbial Physiology
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Author:
Date:
2018-04-20
[Abstract] The human body is colonized by vast communities of microbes, collectively known as microbiota, or microbiome. Although microbes colonize every surface of our bodies that is exposed to the external environment, the biggest collection of microbes colonizing humans and other mammals can be found in the gastrointestinal tract. Given the fact that the human gut is colonized by several hundred microbial species, our group hypothesized that the chemical diversity of this environment should be significant, and that many of the molecules present in that environment would have important signaling roles. Therefore, we devised a protocol to extract these molecules from human feces and test their signaling properties. Potentially bioactive extracts can be tested through addition to culture medium and ...
[摘要] 人体被巨大的微生物群体统称为微生物群体或微生物群体。尽管微生物在我们身体的每一个暴露于外部环境的表面上定殖，但人类和其他哺乳动物中最大量的微生物可以在胃肠道中找到。鉴于人类肠道已被数百种微生物物种繁殖，我们的团队假设这种环境的化学多样性应该是显着的，并且该环境中存在的许多分子将具有重要的信号传导作用。因此，我们制定了一个协议，从人类粪便中提取这些分子并测试其信号特性。可以通过添加培养基并分析细菌生长和基因表达以及其他性质来测试潜在的生物活性提取物。本文描述的方案提供了一种简便且快速的方法，用于使用肠道沙门氏菌作为模型生物体从粪便样品中提取和测试代谢物。该方案还可以适用于从其他基质如培养的哺乳动物细胞，组织，体液和无菌微生物培养物中提取小分子，并且可以针对各种微生物物种测试所得提取物。
【背景】复杂的微生物群落生活在人类和人类身上，将暴露于外部环境的每个表面都定殖。几十年来，这些社区已经获得了几个教派，其中包括正常的植物群，微生物群和最近的微生物群（Sekirov等人，2010年; ...
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| Testing the Effect of UV Radiation on the Survival of Burkholderia glumae
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Author:
Date:
2016-03-05
[Abstract] Burkholderia glumae (B. glumae) is becoming a serious threat in the major rice producing areas of the world. It was reported that Burkholderia spp., including B. glumae, are adapted to a wide range of ecological niches. Different bacterial strains show different levels of UV tolerance which may be due to the presence of different protection mechanisms. Previously we reported that pigment producing strains of B. glumae are more tolerant to UV radiation than non-pigmented strains. Here, we describe the protocol of UV tolerance assay for B. glumae in different exposure times. Using this protocol, we can calculate the survival rate of B. glumae strains, as well as other bacterial species, in exposure to UV radiation.
[摘要] Burkholderia glumae （ B。glumae ）正在成为世界主要稻米生产地区的严重威胁。 据报道，伯克霍尔德菌属包括 B。 腮腺炎，适应广泛的生态位。 不同的细菌菌株显示不同水平的UV耐受性，这可能是由于存在不同的保护机制。 以前，我们报告了颜色产生菌株的B。 腮腺炎比非色素菌株更耐紫外线辐射。 在这里，我们描述了紫外线耐受性测定的方案。 腮腺炎在不同的暴露时间。 使用这个协议，我们可以计算的生存率。 腮腺炎菌株，以及其他细菌物种，暴露于紫外线辐射。
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| In vitro Real-time Measurement of the Intra-bacterial Redox Potential
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Author:
Date:
2015-09-05
[Abstract] All bacteria that live in oxygenated environments have to deal with oxidative stress caused by some form of exogenous or endogenous reactive oxygen species (ROS) (Imlay, 2013). Large quantities of ROS damage DNA, lipids and proteins which can eventually lead to bacterial cell death (Imlay, 2013). In contrast, smaller quantities of ROS can play more sophisticated roles in cellular signalling pathways affecting almost every process in the bacterial cell e.g. metabolism, stress responses, transcription, protein synthesis, etc. Previously, inadequate analytical methods prevented appropriate analysis of the intra-bacterial redox potential. Herein, we describe a method for the measurement of real-time changes to the intra-bacterial redox potential using redox-sensitive GFP ...
[摘要] 所有存在氧合环境中的细菌都必须处理由某种形式的外源性或内源性活性氧（ROS）引起的氧化应激（Imlay，2013）。大量的ROS损伤DNA，脂质和蛋白质，其可以最终导致细菌细胞死亡（Imlay，2013）。相反，较少量的ROS可以在细胞信号传导途径中发挥更复杂的作用，影响几乎细菌细胞中的每一个过程，例如，代谢，应激反应，转录，蛋白质合成等 。以前，不充分的分析方法阻止了细菌内氧化还原电位的适当分析。在本文中，我们描述了使用氧化还原敏感性GFP（roGFP2）测量细菌内氧化还原电位的实时变化的方法（van der Heijden等人，2015）。 roGFP2蛋白被工程改造成含有特定的半胱氨酸残基，其在氧化时形成内部二硫键，导致蛋白构象的轻微改变（Hanson等人，2004）。这种移位导致两种不同的蛋白质同种型在分别在405nm和480nm下激发后具有不同的荧光激发光谱。因此，相应的405/480nm比率可以用作细菌内氧化还原电位的量度。比率量度分析排除了由于roGFP2浓度差异引起的变化，并且由于构象变化是可逆的，因此系统允许测量氧化以及还原条件。在该方案中，我们通过测量鼠伤寒沙门氏菌（鼠伤寒沙门氏菌）内的细菌内氧化还原电位来描述该系统，但该系统可以调整用于其他革兰氏阴性菌。
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