6,142 research outputs found

    Identification of functional cis-regulatory elements by sequential enrichment from a randomized synthetic DNA library

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    BACKGROUND: The identification of endogenous cis-regulatory DNA elements (CREs) responsive to endogenous and environmental cues is important for studying gene regulation and for biotechnological applications but is labor and time intensive. Alternatively, by taking a synthetic biology approach small specific DNA binding sites tailored to the needs of the scientist can be generated and rapidly identified. RESULTS: Here we report a novel approach to identify stimulus-responsive synthetic CREs (SynCREs) from an unbiased random synthetic element (SynE) library. Functional SynCREs were isolated by screening the SynE libray for elements mediating transcriptional activity in plant protoplasts. Responsive elements were chromatin immunoprecipitated by targeting the active Ser-5 phosphorylated RNA polymerase II CTD (Pol II ChIP). Using sequential enrichment, deep sequencing and a bioinformatics pipeline, candidate responsive SynCREs were identified within a pool of constitutively active DNA elements and further validated. These included bonafide biotic/abiotic stress-responsive motifs along with novel SynCREs. We tested several SynCREs in Arabidopsis and confirmed their response to biotic stimuli. CONCLUSIONS: Successful isolation of synthetic stress-responsive elements from our screen illustrates the power of the described methodology. This approach can be applied to any transfectable eukaryotic system since it exploits a universal feature of the eukaryotic Pol II

    Amorphous interface layer in thin graphite films grown on the carbon face of SiC

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    Cross-sectional transmission electron microscopy (TEM) is used to characterize an amorphous layer observed at the interface in graphite and graphene films grown via thermal decomposition of C-face 4H-SiC. The amorphous layer does not to cover the entire interface, but uniform contiguous regions span microns of cross-sectional interface. Annular dark field scanning transmission electron microscopy (ADF-STEM) images and electron energy loss spectroscopy (EELS) demonstrate that the amorphous layer is a carbon-rich composition of Si/C. The amorphous layer is clearly observed in samples grown at 1600{\deg}C for a range of growth pressures in argon, but not at 1500{\deg}C, suggesting a temperature-dependent formation mechanism

    The Effects of Workforce Creativity on Earnings in U.S. Counties

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    This paper examines the effects of local workforce creativity on county-level earnings. Descriptive analysis of the data shows that most of the high-creativity counties in the United States are part of metropolitan areas, and that employee earnings are high in these places. Regression results indicate that, other things being equal, workforce creativity enhances county-level labor earnings. However, the returns to creativity that we found can be confirmed only in the urban context. An extension of the analysis suggests that the creative workforce wage premium may be capturing the effects of "technical workforce creativity" on earnings.creative economy, wages, economic development, Labor and Human Capital,

    Educating Lawyers: Preparation for the Profession of Law

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    Examines the unique aspects and limitations of legal education, as part of a series of reports from the foundation's Preparation for the Professions Program

    Being a Child Today Is Simply Not Easy

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    Issue Editor, Ira Colby and Catherine M. Flaitz\u27s, point of view and summary of the articles in Accountable Communities: Healthier Neighborhoods, Healthier Children
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