48 research outputs found

    Production of Recombinant Human DNA Polymerase Delta in a Bombyx mori Bioreactor

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    Eukaryotic DNA polymerase δ (pol δ) plays a crucial role in chromosomal DNA replication and various DNA repair processes. It is thought to consist of p125, p66 (p68), p50 and p12 subunits. However, rigorous isolation of mammalian pol δ from natural sources has usually yielded two-subunit preparations containing only p125 and p50 polypeptides. While recombinant pol δ isolated from infected insect cells have some problems of consistency in the quality of the preparations, and the yields are much lower. To address these deficiencies, we have constructed recombinant BmNPV baculoviruses using MultiBac system. This method makes the generation of recombinant forms of pol δ containing mutations in any one of the subunits or combinations thereof extremely facile. From about 350 infected larvae, we obtained as much as 4 mg of pol δ four-subunit complex. Highly purified enzyme behaved like the one of native form by rigorous characterization and comparison of its activities on poly(dA)/oligo(dT) template-primer and singly primed M13 DNA, and its homogeneity on FPLC gel filtration. In vitro base excision repair (BER) assays showed that pol δ plays a significant role in uracil-intiated BER and is more likely to mediate LP BER, while the trimer lacking p12 is more likely to mediate SN BER. It seems likely that loss of p12 modulates the rate of SN BER and LP BER during the repair process. Thus, this work provides a simple, fast, reliable and economic way for the large-scale production of human DNA polymerase δ with a high activity and purity, setting up a new platform for our further research on the biochemical properties of pol δ, its regulation and the integration of its functions, and how alterations in pol δ function could contribute to the etiology of human cancer or other diseases that can result from loss of genomic stability

    Measurement of prompt J/ψ pair production in pp collisions at √s = 7 Tev

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    Searches for electroweak production of charginos, neutralinos, and sleptons decaying to leptons and W, Z, and Higgs bosons in pp collisions at 8 TeV

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    Study of hadronic event-shape variables in multijet final states in pp collisions at √s=7 TeV

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    Constraints on parton distribution functions and extraction of the strong coupling constant from the inclusive jet cross section in pp collisions at √s=7 TeV

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    Study of Vector Boson Scattering and Search for New Physics in Events with Two Same-Sign Leptons and Two Jets

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    A study of vector boson scattering in pp collisions at a center-of-mass energy of 8 TeV is presented. The data sample corresponds to an integrated luminosity of 19.4 fb(-1) collected with the CMS detector. Candidate events are selected with exactly two leptons of the same charge, two jets with large rapidity separation and high dijet mass, and moderate missing transverse energy. The signal region is expected to be dominated by electroweak same-sign W-boson pair production. The observation agrees with the standard model prediction. The observed significance is 2.0 standard deviations, where a significance of 3.1 standard deviations is expected based on the standard model. Cross section measurements for (WW +/-)-W-+/- and WZ processes in the fiducial region are reported. Bounds on the structure of quartic vector-boson interactions are given in the framework of dimension-eight effective field theory operators, as well as limits on the production of doubly charged Higgs bosons

    Controllably coated graphene oxide particles with enhanced compatibility with poly(ethylene-co-propylene) thermoplastic elastomer for excellent photo-mechanical actuation capability

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    This paper reports the utilization of the controllable coating via SI-ATRP technique as a promising approach for controlling stimuli-responsive capabilities of graphene oxide (GO) based nanocomposites. Various polymer brushes with controlled molar mass and narrow dispersity were grown from the surface of GO particles. Modification of GO with poly(methyl methacrylate) and poly(n-butyl methacrylate) was proved by transmission electron microscopy, thermogravimetric analysis with online FTIR recording and finally by X-ray photoelectron spectroscopy (XPS). Densities of GO-based materials were investigated and conductivity measurements showed the increase values. XPS and Raman shift was used to confirm the GO particles reduction. A compatibility of the filler with propylene-based elastomer was elucidated by melt rheology. The light-induced actuation capability was investigated on composite samples to show, that polymer hybrid particles based on GO have better compatibility with the polymer matrix and thus their proper dispersibility was significantly improved. In addition the plasticizing effect of the short polymer grafts present on the GO filler surface has the crucial impact on the matrix stiffness and thus the ability of composite material to reversibly respond to the external light stimulation.Scopu
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