4,692 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,

    Effects of fire frequency and the red imported fire ant on native insects in a Louisiana longleaf pine savanna

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    Ants and ground beetles were collected in pitfall traps from a longleaf pine savanna in the early stages of restoration. Insect abundance and species richness were compared among four treatment combinations of two fire frequencies and presence and absence of an exotic ant, Solenopsis invicta, the red imported fire ant. Amdro® was used to suppress S. invicta. Fire treatments consisted of a single fire in a two year period or two fires in a two year period, one applied each year. Fires were applied late in the growing season both years. Ten of the 28 ground beetle species collected were single individuals. Samples were not large enough analyze statistically. Burning a fire-maintained habitat does not typically alter ground beetle populations, but there were not enough data to validate this claim. Twenty-six species of ants were collected from pitfall traps. Six species were exotic. Exotics comprised 23% of the species richness and 98% of the individuals collected. Solenopsis invicta far exceeded other species\u27 abundances and accounted for 95% of total ants collected. Excluding S. invicta, natives and exotics were relatively equal in abundance. Regression analyses of species ranks and abundances were not different among treatments. Species composition among treatments was more similar following treatment applications. After many years of fire suppression, many ant species increased in abundance following the first applied fire. However, the second applied fire did not elicit the same magnitude of response. Soil moisture, soil temperature, and leaf litter dry weights were not correlated with ant abundance. Because there were no differences in native abundance among treatments, it was concluded that Amdro® was not necessary to maintain a balance between S. invicta and native ants. A single applied fire without Amdro® produced the same results with respect to S. invicta and native ant abundance as two fires with Amdro®. The greatest number of exotic ants were collected from treatments with two fires and no Amdro®, suggesting more frequent fires increase exotic ant abundance. Data suggest that native ants are able to coexist with S. invicta and other exotics even when vastly outnumbered by them

    Arthropod dissemination of Discula destructiva : conidia on Cornus florida

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    In 1978, a fungal disease, identified as Discula destructiva Redlin sp. Nov., was discovered killing native flowering dogwoods, Comus florida Link., in the northeastern United States. Since then, the pathogen has spread southward along the Appalachian mountain range to Georgia and Alabama, infecting dogwoods on more than 2.3 million hectares. Known methods of pathogen dissemination include wind and rain. Other possible mechanisms for dispersing fungal conidia involve animal vectors, such as arthropods which may carry conidia externally and/or internally. The objective of this research was to determine if arthropods could disseminate viable D. destructiva\u3c\em\u3e conidia. The adult convergent lady beetle (CLB), Hippodamia convergens\u3c\em\u3e Guerin- Menfeville (Coleoptera: Coccinellidae), was selected as the model insect for this research. In experiments testing for viability of conidia carried externally and internally, CLBs exposed to TN 8 carried viable conidia externally and internally, 100% of the time. All beetles infested with VA 17b carried viable conidia externally and 95% internally. These results were supported by data from time trials in which infested CLBs carried and deposited viable conidia as many as 16 d after exposure to the inoculum. Scanning electron microscopy of CLB body surface showed significantly more conidia were attached to the ventral surface. No difference in conidia densities were observed among body regions on the ventral surface; however, regions within the dorsal surface did have significantly different amounts of conidia. Greenhouse studies to determine if infested CLBs could initiate infection of healthy C. florida\u3c\em\u3e trees produced significant differences when infection by noninoculated CLBs, no CLBs, and inoculated CLBs were compared. Ninety percent of trees treated with inoculated CLBs displayed symptoms of D. destructiva\u3c\em\u3e infection. Scanning electron micrographs of leaves exposed to infested CLBs were observed for locations of deposited conidia. Conidia were deposited around trichomes on the adaxial C. florida leaf surface. Insects are abundant and active organisms that inhabit the forest/urban ecosystems and that, indirectly, may play a role in epidemiology of dogwood anthracnose. Results from this research have indicated that insects can disseminate viable D. destructiva\u3c\em\u3e conidia and may be involved in the rapid movement of the pathogen along the eastern United States throughout C. florida\u27s\u3c\em\u3e habitat range
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