1,264 research outputs found
Rapid antibody selection by mRNA display on a microfluidic chip
In vitro antibody-display technologies are powerful approaches for isolating monoclonal antibodies from recombinant antibody libraries. However, these display techniques require several rounds of affinity selection which is time-consuming. Here, we combined mRNA display with a microfluidic system for in vitro selection and evolution of antibodies and achieved ultrahigh enrichment efficiency of 106- to 108-fold per round. After only one or two rounds of selection, antibodies with high affinity and specificity were obtained from naĂŻve and randomized single-chain Fv libraries of âŒ1012 molecules. Furthermore, we confirmed that not only proteinâprotein (antigenâantibody) interactions, but also proteinâDNA and proteinâdrug interactions were selected with ultrahigh efficiencies. This method will facilitate high-throughput preparation of antibodies and identification of protein interactions in proteomic and therapeutic fields
A Comprehensive Resource of Interacting Protein Regions for Refining Human Transcription Factor Networks
Large-scale data sets of protein-protein interactions (PPIs) are a valuable resource for mapping and analysis of the topological and dynamic features of interactome networks. The currently available large-scale PPI data sets only contain information on interaction partners. The data presented in this study also include the sequences involved in the interactions (i.e., the interacting regions, IRs) suggested to correspond to functional and structural domains. Here we present the first large-scale IR data set obtained using mRNA display for 50 human transcription factors (TFs), including 12 transcription-related proteins. The core data set (966 IRs; 943 PPIs) displays a verification rate of 70%. Analysis of the IR data set revealed the existence of IRs that interact with multiple partners. Furthermore, these IRs were preferentially associated with intrinsic disorder. This finding supports the hypothesis that intrinsically disordered regions play a major role in the dynamics and diversity of TF networks through their ability to structurally adapt to and bind with multiple partners. Accordingly, this domain-based interaction resource represents an important step in refining protein interactions and networks at the domain level and in associating network analysis with biological structure and function
Particle velocity in noncommutative space-time
We investigate a particle velocity in the -Minkowski space-time,
which is one of the realization of a noncommutative space-time. We emphasize
that arrival time analyses by high-energy -rays or neutrinos, which
have been considered as powerful tools to restrict the violation of Lorentz
invariance, are not effective to detect space-time noncommutativity. In
contrast with these examples, we point out a possibility that {\it low-energy
massive particles} play an important role to detect it.Comment: 16 pages, corrected some mistake
Use of cDNA Tiling Arrays for Identifying Protein Interactions Selected by In Vitro Display Technologies
In vitro display technologies such as mRNA display are powerful screening tools for protein interaction analysis, but the final cloning and sequencing processes represent a bottleneck, resulting in many false negatives. Here we describe an application of tiling array technology to identify specifically binding proteins selected with the in vitro virus (IVV) mRNA display technology. We constructed transcription-factor tiling (TFT) arrays containing âŒ1,600 open reading frame sequences of known and predicted mouse transcription-regulatory factors (334,372 oligonucleotides, 50-mer in length) to analyze cDNA fragments from mRNA-display screening for Jun-associated proteins. The use of the TFT arrays greatly increased the coverage of known Jun-interactors to 28% (from 14% with the cloning and sequencing approach), without reducing the accuracy (âŒ75%). This method could detect even targets with extremely low expression levels (less than a single mRNA copy per cell in whole brain tissue). This highly sensitive and reliable method should be useful for high-throughput protein interaction analysis on a genome-wide scale
Ligand-Receptor Interactions
The formation and dissociation of specific noncovalent interactions between a
variety of macromolecules play a crucial role in the function of biological
systems. During the last few years, three main lines of research led to a
dramatic improvement of our understanding of these important phenomena. First,
combination of genetic engineering and X ray cristallography made available a
simultaneous knowledg of the precise structure and affinity of series or
related ligand-receptor systems differing by a few well-defined atoms. Second,
improvement of computer power and simulation techniques allowed extended
exploration of the interaction of realistic macromolecules. Third, simultaneous
development of a variety of techniques based on atomic force microscopy,
hydrodynamic flow, biomembrane probes, optical tweezers, magnetic fields or
flexible transducers yielded direct experimental information of the behavior of
single ligand receptor bonds. At the same time, investigation of well defined
cellular models raised the interest of biologists to the kinetic and mechanical
properties of cell membrane receptors. The aim of this review is to give a
description of these advances that benefitted from a largely multidisciplinar
approach
Emulsion sheet doublets as interface trackers for the OPERA experiment
New methods for efficient and unambiguous interconnection between electronic
counters and target units based on nuclear photographic emulsion films have
been developed. The application to the OPERA experiment, that aims at detecting
oscillations between mu neutrino and tau neutrino in the CNGS neutrino beam, is
reported in this paper. In order to reduce background due to latent tracks
collected before installation in the detector, on-site large-scale treatments
of the emulsions ("refreshing") have been applied. Changeable Sheet (CSd)
packages, each made of a doublet of emulsion films, have been designed,
assembled and coupled to the OPERA target units ("ECC bricks"). A device has
been built to print X-ray spots for accurate interconnection both within the
CSd and between the CSd and the related ECC brick. Sample emulsion films have
been extensively scanned with state-of-the-art automated optical microscopes.
Efficient track-matching and powerful background rejection have been achieved
in tests with electronically tagged penetrating muons. Further improvement of
in-doublet film alignment was obtained by matching the pattern of low-energy
electron tracks. The commissioning of the overall OPERA alignment procedure is
in progress.Comment: 19 pages, 19 figure
Search for DCC in 158A GeV Pb+Pb Collisions
A detailed analysis of the phase space distributions of charged particles and
photons have been carried out using two independent methods. The results
indicate the presence of nonstatistical fluctuations in localized regions of
phase space.Comment: Talk at the PANIC99 Conference, June 9-16, 199
Measurement and comparison of individual external doses of high-school students living in Japan, France, Poland and Belarus -- the "D-shuttle" project --
Twelve high schools in Japan (of which six are in Fukushima Prefecture), four
in France, eight in Poland and two in Belarus cooperated in the measurement and
comparison of individual external doses in 2014. In total 216 high-school
students and teachers participated in the study. Each participant wore an
electronic personal dosimeter "D-shuttle" for two weeks, and kept a journal of
his/her whereabouts and activities. The distributions of annual external doses
estimated for each region overlap with each other, demonstrating that the
personal external individual doses in locations where residence is currently
allowed in Fukushima Prefecture and in Belarus are well within the range of
estimated annual doses due to the background radiation level of other
regions/countries
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