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Impact of nanoscale topography on genomics and proteomics of adherent bacteria

机译:纳米形貌对粘附细菌的基因组学和蛋白质组学的影响

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Bacterial adhesion onto inorganicanoengineered surfaces is a key issue in biotechnology and medicine, because it is one of the first necessary steps to determine a general pathogenic event. Understanding the molecular mechanisms of bacteria-surface interaction represents a milestone for planning a new generation of devices with unanimously certified antibacterial characteristics. Here, we show how highly controlled nanostructured substrates impact the bacterial behavior in terms of morphological, genomic, and proteomic response. We observed by atomic force microscopy (AFM) and scanning electron microscopy (SEM) that type-1 fimbriae typically disappear in Escherichia coli adherent onto nanostructured substrates, as opposed to bacteria onto reference glass or flat gold surfaces. A genetic variation of the fimbrial operon regulation was consistently identified by real time qPCR in bacteria interacting with the nanorough substrates. To gain a deeper insight into the molecular basis of the interaction mechanisms, we explored the entire proteomic profile of E. coli by 2D-DIGE, finding significant changes in the bacteria adherent onto the nanorough substrates, such as regulations of proteins involved in stress processes and defense mechanisms. We thus demonstrated that a pure physical stimulus, that is, a nanoscale variation of surface topography, may play per se a significant role in determining the morphological, genetic, and proteomic profile of bacteria. These data suggest that in depth investigations of the molecular processes of microorganisms adhering to surfaces are of great importance for the design of innovative biomaterials with active biological functionalities.
机译:在无机/纳米工程表面上的细菌粘附是生物技术和医学中的关键问题,因为它是确定一般致病事件的首要必要步骤之一。了解细菌与表面相互作用的分子机制是规划具有一致认证抗菌特性的新一代设备的里程碑。在这里,我们展示了高度受控的纳米结构底物如何在形态,基因组和蛋白质组学响应方面影响细菌行为。我们通过原子力显微镜(AFM)和扫描电子显微镜(SEM)观察到,与粘附在参考玻璃或平坦金表面上的细菌相反,类型1菌毛通常在粘附在纳米结构基质上的大肠杆菌中消失。通过实时qPCR在与纳米粗糙底物相互作用的细菌中一致地确定了纤维操纵子调控的遗传变异。为了更深入地了解相互作用机制的分子基础,我们通过2D-DIGE探索了大肠杆菌的整个蛋白质组学特征,发现粘附在纳米粗糙基质上的细菌发生了显着变化,例如应激过程中涉及的蛋白质调控和防御机制。因此,我们证明了纯物理刺激,即表面形貌的纳米级变化,本身可能在确定细菌的形态,遗传和蛋白质组学方面起着重要作用。这些数据表明,深入研究粘附在表面上的微生物的分子过程对于设计具有活性生物功能的创新生物材料非常重要。

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