20 research outputs found

    Single Bead Affinity Detection (SINBAD) for the Analysis of Protein-Protein Interactions

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    We present a miniaturized pull-down method for the detection of protein-protein interactions using standard affinity chromatography reagents. Binding events between different proteins, which are color-coded with quantum dots (QDs), are visualized on single affinity chromatography beads by fluorescence microscopy. The use of QDs for single molecule detection allows the simultaneous analysis of multiple protein-protein binding events and reduces the amount of time and material needed to perform a pull-down experiment

    Finishing the euchromatic sequence of the human genome

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    The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∌99% of the euchromatic genome and is accurate to an error rate of ∌1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

    Cell cycle dependent differences in nuclear pore complex assembly

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    In metazoa, nuclear pore complexes (NPCs) assemble from disassembled precursors into a reforming nuclear envelope (NE) at the end of mitosis, and into growing intact NEs during interphase. Whether there are differences in the mechanism of NPC assembly in these two scenarios is a long standing question in the field that has not been addressed. Experiments described in this dissertation, show that ELYS, a nucleoporin critical for the recruitment of the essential Nup107/160 complex to chromatin, is crucial for NPC assembly at the end of mitosis, but is not required in interphase. Conversely, the transmembrane nucleoporin POM121 is critical for the incorporation of the Nup107/160 complex into new assembly sites specifically during interphase and plays a role in fusing the two leaflets of the NE. We show that in contrast to post-mitosis, where the Nup107/160 complex is targeted to chromatin via ELYS, during interphase this NPC sub-complex assembles at sites of forming pores. These results indicate that, in organisms with open mitosis, NPCs assemble by two distinct mechanisms to accommodate cell cycle-dependent differences in NE topolog

    NUCLEAR ENVELOPE AND GENOME INTERACTIONS IN CELL FATE

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    The eukaryotic cell nucleus houses an organism’s genome and is the location within the cell, where all signaling-induced and development-driven gene expression programs are ultimately specified. The genome is enclosed and separated from the cytoplasm by the nuclear envelope (NE), a double-lipid membrane bilayer, which contains a large variety of trans-membrane and associated protein complexes. In recent years, research regarding multiple aspects of the cell nucleus points to a highly dynamic and coordinated concert of efforts between chromatin and the NE in regulation of gene expression. Details of how this concert is orchestrated and how it directs cell differentiation and disease are coming to light at a rapid pace. Here we review existing and emerging concepts of how interactions between the genome and the NE may contribute to tissue-specific gene expression programs to determine cell fate

    Taxonomic history and review of the Förster genera of Platygastridae (Hymenoptera: Platygastroidea)

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    Platygastridae is a Ê»dark taxonÊŒ, with many genera and species in dire need of professional attention. The taxonomic impediment is especially severe in the Palearctic Platygastrinae due to the abundance of names with vague concepts. Historical descriptions and their associated type material must be examined and clarified before further revisionary work can occur. Arnold Förster described 18 genera of Platygastridae, most of which represent distinct and recognizable lineages. The present study reviews their taxonomic history, providing diagnostic remarks, English translations, and illustrations of important specimens from the Förster collection in the Natural History Museum Vienna. The collection also includes original exemplar specimens of European species whose types have been lost. Neotypes and lectotypes are designated from this material to improve nomenclatural stability in the group. Neotypes are designated for Amblyaspis forticornis (Nees, 1834), Isocybus grandis (Nees, 1834), Platygaster striolata Nees, 1834, and Trichacis tristis (Nees, 1834). Lectotypes are designated for Leptacis spinigera (Nees, 1834) comb. nov. and for Platygaster corvina Förster, 1861, with Platygaster henkvlugi Buhl, 1996 treated as a junior synonym. Platygaster mutica Nees, 1834 stat. rev., nomen dubium, is transferred from Synopeas

    A fortuitous find: a unique haplotype of Ooencyrtus nezarae Ishii (Encyrtidae: Encyrtinae) discovered in Florida

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    The adventive arrival of biological control agents circumvents the regulatory process by introducing exotic species to control invasive pests and is generally followed by post hoc risk evaluation. The bean plataspid, Megacopta cribraria (Fabricius) (Hemiptera: Plataspidae), is an invasive pest of leguminous crops in the south-eastern United States that was eventually followed by two parasitoid wasps from its range in the eastern hemisphere, Paratelenomus saccharalis (Dodd) (Scelionidae) and Ooencyrtus nezarae Ishii (Encyrtidae). In North Central Florida, sentinel egg masses, intended to capture Paratelenomus saccharalis, instead yielded Ooencyrtus nezarae, which was previously known only from Alabama (Ademokoya et al. 2018). Two generations of O. nezarae were subsequently reared in the laboratory. COI sequences from the Florida population of O. nezarae differed by 1.3% from the Alabama population and the presence of a different haplotype suggests the possibility of a separate introduction. Laboratory parasitism rates, sex ratios, morphology, molecular diagnosis and implications for agriculture are discussed

    Single Bead Affinity Detection (SINBAD) for the Analysis of Protein-Protein Interactions

    No full text
    International audienceWe present a miniaturized pull-down method for the detection of protein-protein interactions using standard affinity chromatography reagents. Binding events between different proteins, which are color-coded with quantum dots (QDs), are visualized on single affinity chromatography beads by fluorescence microscopy. The use of QDs for single molecule detection allows the simultaneous analysis of multiple protein-protein binding events and reduces the amount of time and material needed to perform a pull-down experiment

    A fortuitous find: a unique haplotype of Ooencyrtus nezarae Ishii (Encyrtidae: Encyrtinae) discovered in Florida

    No full text
    The adventive arrival of biological control agents circumvents the regulatory process by introducing exotic species to control invasive pests and is generally followed by post hoc risk evaluation. The bean plataspid, Megacopta cribraria (Fabricius) (Hemiptera: Plataspidae), is an invasive pest of leguminous crops in the south-eastern United States that was eventually followed by two parasitoid wasps from its range in the eastern hemisphere, Paratelenomus saccharalis (Dodd) (Scelionidae) and Ooencyrtus nezarae Ishii (Encyrtidae). In North Central Florida, sentinel egg masses, intended to capture Paratelenomus saccharalis, instead yielded Ooencyrtus nezarae, which was previously known only from Alabama (Ademokoya et al. 2018). Two generations of O. nezarae were subsequently reared in the laboratory. COI sequences from the Florida population of O. nezarae differed by 1.3% from the Alabama population and the presence of a different haplotype suggests the possibility of a separate introduction. Laboratory parasitism rates, sex ratios, morphology, molecular diagnosis and implications for agriculture are discussed
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