115 research outputs found

    Methods for expression of recombinant proteins using a Pichia pastoris cell-free system

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    Cell‐free protein synthesis is a powerful tool for engineering biology and has been utilized in many diverse applications, from biosensing and protein prototyping to biomanufacturing and the design of metabolic pathways. By exploiting host cellular machinery decoupled from cellular growth, proteins can be produced in vitro both on demand and rapidly. Eukaryotic cell‐free platforms are often neglected due to perceived complexity and low yields relative to their prokaryotic counterparts, despite providing a number of advantageous properties. The yeast Pichia pastoris (also known as Komagataella phaffii) is a particularly attractive eukaryotic host from which to generate cell‐free extracts, due to its ability to grow to high cell densities with high volumetric productivity, genetic tractability for strain engineering, and ability to perform post‐translational modifications. Here, we describe methods for conducting cell‐free protein synthesis using P. pastoris as the host, from preparing the cell lysates to protocols for both coupled and linked transcription‐translation reactions. By providing these methodologies, we hope to encourage the adoption of the platform by new and experienced users alike

    Desinfestação de explantes radiculares de bacurizeiro.

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    Este trabalho teve como objetivo avaliar a desinfestação de explantes radiculares de bacurizeiro para o seu estabelecimento in vitro

    Overview of NASA FINESSE (Field Investigations to Enable Solar System Science and Exploration) Science and Exploration Results

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    NASA's FINESSE (Field Investigations to Enable Solar System Science and Exploration) project is focused on a science and exploration field-based research program to generate strategic knowledge in preparation for human and robotic exploration of other planetary bodies including our moon, Mars' moons Phobos and Deimos, and near-Earth asteroids. Scientific study focuses on planetary volcanism (e.g., the formation of volcanoes, evolution of magma chambers and the formation of multiple lava flow types, as well as the evolution and entrapment of volatile chemicals) and impact cratering (impact rock modification, cratering mechanics, and the chronologic record). FINESSE conducts multiple terrestrial field campaigns (Craters of the Moon National Monument and Preserve in Idaho for volcanics, and West Clearwater Impact Structure in Canada for impact studies) to study such features as analogs relevant to our moon, Phobos, Deimos, and asteroids. Here we present the science and exploration results from two deployments to Idaho (2014, 2015) and our first deployment to Canada (2014). FINESSE was selected as a research team by NASA's Solar System Exploration Research Virtual Institute (SSERVI). SSERVI is a joint effort by NASA's Science Mission Directorate (SMD) and Human Exploration and Operations Mission Directorate (HEOMD)

    Deep-Sea Exploration of the US Gulf of Mexico with NOAA Ship Okeanos Explorer

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    Oceanography articles are licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution, and reproduction in any medium or format as long as users cite the materials appropriately (e.g., authors, Oceanography, volume number, issue number, page number[s], figure number[s], and DOI for the article), provide a link to the Creative Commons license, and indicate the changes that were made to the original content

    Direct 2D spatial coherence determination using the Fourier analysis method Multi parameter characterization of the P04 beamline at PETRA III

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    We present a systematic 2D spatial-coherence analysis of the soft-X-ray beamline P04 at PETRA III for various beamline configurations. The influence of two different beam-defining apertures on the spatial coherence properties of the beam is discussed and optimal conditions for coherence-based experiments are found. A significant degradation of the spatial coherence in the vertical direction has been measured and sources of this degradation are identified and discussed. The Fourier-analysis method, which gives fast and simple access to the 2D spatial coherence function of the X-ray beam, is used for the experiment. Here, we exploit the charge scattering of a disordered nanodot sample allowing the use of arbitrary X-ray photon energies with this method

    Friction reduction and zero wear for 52100 bearing steel by high‐dose implantation of carbon

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    Ion implantation of carbon in the AISI 52100 bearing steel yields a distinct reduction in friction and wear. This improvement is strongly dependent on the implanted fluence. The coefficient of friction decreases from 0.6 to 0.2 for doses >1×1018 cm-2 (energy 100 keV) and a wear reduction to nearly ‘‘zero wear’’ was obtainable even under severe wear conditions. The counterpart (unimplanted AISI 52100 steel ball) shows a similar behavior, which demonstrates that the tribological system is totally changed. Mössbauer spectroscopy and x-ray diffraction revealed that hexagonal ¿-carbide is formed on implantation. On the other hand, Rutherford backscattering spectrometry shows that for high doses a large fraction of the implanted carbon is not contained in this carbide
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