185 research outputs found

    Rearranging the centromere of the human Y chromosome with Ļ†C31 integrase

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    We have investigated the ability of the integrase from the Streptomyces Ļ†C31 ā€˜phage to either delete or invert 1 Mb of DNA around the centromere of the human Y chromosome in chicken DT40 hybrid somatic cells. Reciprocal and conservative site-specific recombination was observed in 54% of cells expressing the integrase. The sites failed to recombine in the remaining cells because the sites had been damaged. The sequences of the damaged sites indicated that the damage arose as a result of repair of recombination intermediates by host cell pathways. The liability of recombination intermediates to damage is consistent with what is known about the mechanism of serine recombinase reactions. The structures of the products of the chromosome rearrangements were consistent with the published sequence of the Y chromosome indicating that the assembly of the highly repeated region between the sites is accurate to a resolution of about 50 kb. Mini-chromosomes lacking a centromere were not recovered which also suggested that neo-centromere formation occurs infrequently in vertebrate somatic cells. No ectopic recombination was observed between a Ļ†C31 integrase attB site and the chicken genome

    Use of the gLite-WMS in CMS for production and analysis

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    The CMS experiment at LHC started using the Resource Broker (by the EDG and LCG projects) to submit Monte Carlo production and analysis jobs to distributed computing resources of the WLCG infrastructure over 6 years ago. Since 2006 the gLite Workload Management System (WMS) and Logging \& Bookkeeping (LB) are used. The interaction with the gLite-WMS/LB happens through the CMS production and analysis frameworks, respectively ProdAgent and CRAB, through a common component, BOSSLite. The important improvements recently made in the gLite-WMS/LB as well as in the CMS tools and the intrinsic independence of different WMS/LB instances allow CMS to reach the stability and scalability needed for LHC operations. In particular the use of a multi-threaded approach in BOSSLite allowed to increase the scalability of the systems significantly. In this work we present the operational set up of CMS production and analysis based on the gLite-WMS and the performances obtained in the past data challenges and in the daily Monte Carlo productions and user analysis usage in the experiment

    RMDAP: A Versatile, Ready-To-Use Toolbox for Multigene Genetic Transformation

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    Background: The use of transgenes to improve complex traits in crops has challenged current genetic transformation technology for multigene transfer. Therefore, a multigene transformation strategy for use in plant molecular biology and plant genetic breeding is thus needed. Methodology/Principal Findings: Here we describe a versatile, ready-to-use multigene genetic transformation method, named the Recombination-assisted Multifunctional DNA Assembly Platform (RMDAP), which combines many of the useful features of existing plant transformation systems. This platform incorporates three widely-used recombination systems, namely, Gateway technology, in vivo Cre/loxP and recombineering into a highly efficient and reliable approach for gene assembly. RMDAP proposes a strategy for gene stacking and contains a wide range of flexible, modular vectors offering a series of functionally validated genetic elements to manipulate transgene overexpression or gene silencing involved in a metabolic pathway. In particular, the ability to construct a multigene marker-free vector is another attractive feature. The built-in flexibility of original vectors has greatly increased the expansibility and applicability of the system. A proof-ofprinciple experiment was confirmed by successfully transferring several heterologous genes into the plant genome. Conclusions/Significance: This platform is a ready-to-use toolbox for full exploitation of the potential for coordinate regulation of metabolic pathways and molecular breeding, and will eventually achieve the aim of what we call ā€˜ā€˜one-sto

    Gender and Management: new directions in research and continuing patterns in practice

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    This is the authorā€™s version of the following article. The definitive version is available at www.interscience.wiley.com:Adelina Broadbridge and Jeff Hearn, Gender and management: New directions in research and continuing patterns in practice, 2008, British Journal of Management, (19), s1, 38-49. http://dx.doi.org/10.1111/j.1467-8551.2008.00570.xCopyright: British Academy of Management, Blackwell Publishing Ltdhttp://www.blackwellpublishing.com

    Recombinase technology: applications and possibilities

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    The use of recombinases for genomic engineering is no longer a new technology. In fact, this technology has entered its third decade since the initial discovery that recombinases function in heterologous systems (Sauer in Mol Cell Biol 7(6):2087ā€“2096, 1987). The random insertion of a transgene into a plant genome by traditional methods generates unpredictable expression patterns. This feature of transgenesis makes screening for functional lines with predictable expression labor intensive and time consuming. Furthermore, an antibiotic resistance gene is often left in the final product and the potential escape of such resistance markers into the environment and their potential consumption raises consumer concern. The use of site-specific recombination technology in plant genome manipulation has been demonstrated to effectively resolve complex transgene insertions to single copy, remove unwanted DNA, and precisely insert DNA into known genomic target sites. Recombinases have also been demonstrated capable of site-specific recombination within non-nuclear targets, such as the plastid genome of tobacco. Here, we review multiple uses of site-specific recombination and their application toward plant genomic engineering. We also provide alternative strategies for the combined use of multiple site-specific recombinase systems for genome engineering to precisely insert transgenes into a pre-determined locus, and removal of unwanted selectable marker genes

    Gendering the careers of young professionals: some early findings from a longitudinal study. in Organizing/theorizing: developments in organization theory and practice

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    Wonders whether companies actually have employees best interests at heart across physical, mental and spiritual spheres. Posits that most organizations ignore their workforce ā€“ not even, in many cases, describing workers as assets! Describes many studies to back up this claim in theis work based on the 2002 Employment Research Unit Annual Conference, in Cardiff, Wales
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