1,374 research outputs found

    Identification of a gene encoding GMP synthetase from a Neurospora crassa cDNA library by bacterial complementation

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    We report the isolation and identification of a gene encoding GMP synthetase from a Neurospora crassa cDNA library. Phage infection of the purine-requiring Escherichia coli strain SØ3834 using the NO3- induced cDNA phage library from the Fungal Genetics Stock Center resulted in colonies able to grow on minimal media with no added purine source. A plasmid, termed pGMPS1, was isolated from one of these colonies and shown to reproducibly support growth of strain SØ3834 in the absence of purines in the media. Identification of this gene as one encoding GMP synthetase is confirmed by DNA sequencing and comparison to the known guaA gene from yeast

    Aphids

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    Methods for Detecting and Quantifying Viable Bacterial Endo-Spores

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    Methods and systems for detecting viable bacterial endospores in a sample and related methods to quantify viable bacterial endospores in a sample

    Toolbox for analyzing finite two-state trajectories

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    In many experiments, the aim is to deduce an underlying multi-substate on-off kinetic scheme (KS) from the statistical properties of a two-state trajectory. However, the mapping of a KS into a two-state trajectory leads to the loss of information about the KS, and so, in many cases, more than one KS can be associated with the data. We recently showed that the optimal way to solve this problem is to use canonical forms of reduced dimensions (RD). RD forms are on-off networks with connections only between substates of different states, where the connections can have non-exponential waiting time probability density functions (WT-PDFs). In theory, only a single RD form can be associated with the data. To utilize RD forms in the analysis of the data, a RD form should be associated with the data. Here, we give a toolbox for building a RD form from a finite two-state trajectory. The methods in the toolbox are based on known statistical methods in data analysis, combined with statistical methods and numerical algorithms designed specifically for the current problem. Our toolbox is self-contained - it builds a mechanism based only on the information it extracts from the data, and its implementation on the data is fast (analyzing a 10^6 cycle trajectory from a thirty-parameter mechanism takes a couple of hours on a PC with a 2.66 GHz processor). The toolbox is automated and is freely available for academic research upon electronic request

    A LEKID-based CMB instrument design for large-scale observations in Greenland

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    We present the results of a feasibility study, which examined deployment of a ground-based millimeter-wave polarimeter, tailored for observing the cosmic microwave background (CMB), to Isi Station in Greenland. The instrument for this study is based on lumped-element kinetic inductance detectors (LEKIDs) and an F/2.4 catoptric, crossed-Dragone telescope with a 500 mm aperture. The telescope is mounted inside the receiver and cooled to < 4<\,4 K by a closed-cycle 4^4He refrigerator to reduce background loading on the detectors. Linearly polarized signals from the sky are modulated with a metal-mesh half-wave plate that is rotated at the aperture stop of the telescope with a hollow-shaft motor based on a superconducting magnetic bearing. The modular detector array design includes at least 2300 LEKIDs, and it can be configured for spectral bands centered on 150~GHz or greater. Our study considered configurations for observing in spectral bands centered on 150, 210 and 267~GHz. The entire polarimeter is mounted on a commercial precision rotary air bearing, which allows fast azimuth scan speeds with negligible vibration and mechanical wear over time. A slip ring provides power to the instrument, enabling circular scans (360 degrees of continuous rotation). This mount, when combined with sky rotation and the latitude of the observation site, produces a hypotrochoid scan pattern, which yields excellent cross-linking and enables 34\% of the sky to be observed using a range of constant elevation scans. This scan pattern and sky coverage combined with the beam size (15~arcmin at 150~GHz) makes the instrument sensitive to 5<ℓ<10005 < \ell < 1000 in the angular power spectra

    The Detector System for the Stratospheric Kinetic Inductance Polarimeter (SKIP)

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    The Stratospheric Kinetic Inductance Polarimeter (SKIP) is a proposed balloon-borne experiment designed to study the cosmic microwave background, the cosmic infrared background and Galactic dust emission by observing 1133 square degrees of sky in the Northern Hemisphere with launches from Kiruna, Sweden. The instrument contains 2317 single-polarization, horn-coupled, aluminum lumped-element kinetic inductance detectors (LEKID). The LEKIDs will be maintained at 100 mK with an adiabatic demagnetization refrigerator. The polarimeter operates in two configurations, one sensitive to a spectral band centered on 150 GHz and the other sensitive to 260 and 350 GHz bands. The detector readout system is based on the ROACH-1 board, and the detectors will be biased below 300 MHz. The detector array is fed by an F/2.4 crossed-Dragone telescope with a 500 mm aperture yielding a 15 arcmin FWHM beam at 150 GHz. To minimize detector loading and maximize sensitivity, the entire optical system will be cooled to 1 K. Linearly polarized sky signals will be modulated with a metal-mesh half-wave plate that is mounted at the telescope aperture and rotated by a superconducting magnetic bearing. The observation program consists of at least two, five-day flights beginning with the 150 GHz observations.Comment: J Low Temp Phys DOI 10.1007/s10909-013-1014-3 The final publication is available at link.springer.co

    Human HERC5 restricts an early stage of HIV-1 assembly by a mechanism correlating with the ISGylation of Gag

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    <p>Abstract</p> <p>Background</p> <p>The identification and characterization of several interferon (IFN)-induced cellular HIV-1 restriction factors, defined as host cellular proteins or factors that restrict or inhibit the HIV-1 life cycle, have provided insight into the IFN response towards HIV-1 infection and identified new therapeutic targets for HIV-1 infection. To further characterize the mechanism underlying restriction of the late stages of HIV-1 replication, we assessed the ability of IFNbeta-induced genes to restrict HIV-1 Gag particle production and have identified a potentially novel host factor called HECT domain and RCC1-like domain-containing protein 5 (HERC5) that blocks a unique late stage of the HIV-1 life cycle.</p> <p>Results</p> <p>HERC5 inhibited the replication of HIV-1 over multiple rounds of infection and was found to target a late stage of HIV-1 particle production. The E3 ligase activity of HERC5 was required for blocking HIV-1 Gag particle production and correlated with the post-translational modification of Gag with ISG15. HERC5 interacted with HIV-1 Gag and did not alter trafficking of HIV-1 Gag to the plasma membrane. Electron microscopy revealed that the assembly of HIV-1 Gag particles was arrested at the plasma membrane, at an early stage of assembly. The mechanism of HERC5-induced restriction of HIV-1 particle production is distinct from the mechanism underlying HIV-1 restriction by the expression of ISG15 alone, which acts at a later step in particle release. Moreover, HERC5 restricted murine leukemia virus (MLV) Gag particle production, showing that HERC5 is effective in restricting Gag particle production of an evolutionarily divergent retrovirus.</p> <p>Conclusions</p> <p>HERC5 represents a potential new host factor that blocks an early stage of retroviral Gag particle assembly. With no apparent HIV-1 protein that directly counteracts it, HERC5 may represent a new candidate for HIV/AIDS therapy.</p
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