WorldWideScience

Sample records for mega plasmid resident

  1. Cereulide synthetase gene cluster from emetic Bacillus cereus: Structure and location on a mega virulence plasmid related to Bacillus anthracis toxin plasmid pXO1

    Wagner Martin

    2006-03-01

    Full Text Available Abstract Background Cereulide, a depsipeptide structurally related to valinomycin, is responsible for the emetic type of gastrointestinal disease caused by Bacillus cereus. Recently, it has been shown that this toxin is produced by a nonribosomal peptide synthetase (NRPS, but its exact genetic organization and biochemical synthesis is unknown. Results The complete sequence of the cereulide synthetase (ces gene cluster, which encodes the enzymatic machinery required for the biosynthesis of cereulide, was dissected. The 24 kb ces gene cluster comprises 7 CDSs and includes, besides the typical NRPS genes like a phosphopantetheinyl transferase and two CDSs encoding enzyme modules for the activation and incorporation of monomers in the growing peptide chain, a CDS encoding a putative hydrolase in the upstream region and an ABC transporter in the downstream part. The enzyme modules responsible for incorporation of the hydroxyl acids showed an unusual structure while the modules responsible for the activation of the amino acids Ala and Val showed the typical domain organization of NRPS. The ces gene locus is flanked by genetic regions with high homology to virulence plasmids of B. cereus, Bacillus thuringiensis and Bacillus anthracis. PFGE and Southern hybridization showed that the ces genes are restricted to emetic B. cereus and indeed located on a 208 kb megaplasmid, which has high similarities to pXO1-like plasmids. Conclusion The ces gene cluster that is located on a pXO1-like virulence plasmid represents, beside the insecticidal and the anthrax toxins, a third type of B. cereus group toxins encoded on megaplasmids. The ces genes are restricted to emetic toxin producers, but pXO1-like plasmids are also present in emetic-like strains. These data might indicate the presence of an ancient plasmid in B. cereus which has acquired different virulence genes over time. Due to the unusual structure of the hydroxyl acid incorporating enzyme modules of Ces

  2. Resident enhanced repair: novel repair process action on plasmid DNA transformed into Escherichia coli K-12

    The survival of UV-irradiated DNA of plasmid NTP16 was monitored after its transformation into recipient cells containing an essentially homologous undamaged plasmid, pLV9. The presence of pLV9 resulted in a substantial increase in the fraction of damaged NTP16 molecules which survived in the recipient cells. This enhanced survival requires the host uvrA+ and uvrB+ gene products, but not the host recA+ gene product. The requirement for both homologous DNA and the uvrA+ gene products suggests that a novel repair process may act on plasmid DNA. Possible mechanisms for this process are considered

  3. Examination of residence time and its relevance to water quality within a coastal mega-structure: The Palm Jumeirah Lagoon

    Cavalcante, Geórgenes H.; Kjerfve, Björn; Feary, David A.

    2012-10-01

    SummaryA numerical modeling study was carried out to compute average residence time in the semi-enclosed lagoon formed by the man-made island Palm Jumeirah (Dubai, United Arab Emirates), termed Palm Jumeirah Lagoon (PJL). The PJL encompasses a main island axis with 17 'fronds' radiating from this axis, all encapsulated within a semi-circular breakwater system. A coupled hydrodynamic and solute transport model was developed for the waters of the PJL, based on depth-integrated conservation equations. Numerical model predictions were then verified against a set of field-measured hydrodynamic data. Model-predicted water elevations and velocities were in good agreement with field measurements. Residence times for this tidal dominated system were investigated through numerical experiments using a conservative tracer as a surrogate. The results indicated that average residence time varied spatially throughout the PJL depending on tidal flushing. Average residence time was unequally distributed throughout the PJL, with the eastern side showing higher flushing times than the western side. In addition, there were also differences between sections of the PJL in average residence time of a tracer: between frond tips and the surrounding breakwater the tracer was reduced to 30-40% of its original value after approximately 1 week, while a tracer placed between the fronds was reduced to 30-40% of its value after 20 days. The findings of this research provide vital information for understanding the water transport process in this man-made lagoon, and will be important in assessing the potential impact on coastal water quality conditions in coastal developments within the Middle East.

  4. Interactions between octopine and nopaline plasmids in Agrobacterium tumefaciens.

    Hooykaas, P J; Den Dulk-Ras, H; Ooms, G.; Schilperoort, R.A.

    1980-01-01

    Transfer of octopine Ti plasmids to strains already carrying an octopine Ti plasmid was found to occur at the same (high) frequency as transfer to Ti plasmid lacking recipients, showing that resident Ti plasmids do not exhibit entry exclusion towards incoming Ti plasmids. The resident octopine Ti plasmid was lost by the recipient after the entrance of the incoming Ti plasmid, which is indicative of the incompatibility between the Ti plasmids. Octopine Ti plasmids were found to become establis...

  5. Plasmid-mediated gene transfer between insect-resident bacteria, Enterobacter cloacae, and plant-epiphytic bacteria, Erwinia herbicola, in guts of silkworm larvae.

    Watanabe, K; Sato, M

    1998-11-01

    Five strains of Enterobacter cloacae isolated from several species of plants and insects were able to grow in the guts of silkworm larvae. A much larger population of Ent. cloacae strains was detected in the insect guts and feces collected 3 and 6 days than in samples collected 1 day after feeding artificial diets contaminating these bacteria. Furthermore, insect-origin strains of Ent. cloacae were mated with a donor strain, epiphytic Erwinia herbicola, harboring RSF1010 and pBPW1::Tn7 plasmids in the insect guts by introducing these bacteria through separate artificial diets administered at different times. A number of transconjugants, Ent. cloacae strains which had acquired RSF1010 plasmid, were detected from guts and fecal samples at transfer frequencies of 10(-2) to 10(-3) per recipient. Thus, gene transfer between epiphytic Er. herbicola and insect-resident Ent. cloacae strains in the insect guts was confirmed. These findings may provide significant information about the role of "in insecta mating" in the evolution of these bacteria. PMID:9767717

  6. Computerized mega code recording.

    Burt, T W; Bock, H C

    1988-04-01

    A system has been developed to facilitate recording of advanced cardiac life support mega code testing scenarios. By scanning a paper "keyboard" using a bar code wand attached to a portable microcomputer, the person assigned to record the scenario can easily generate an accurate, complete, timed, and typewritten record of the given situations and the obtained responses. PMID:3354937

  7. Mega Key Authentication Mechanism

    Kloss, Guy

    2016-01-01

    For secure communication it is not just sufficient to use strong cryptography with good and strong keys, but to actually have the assurance, that the keys in use for it are authentic and from the contact one is expecting to communicate with. Without that, it is possible to be subject to impersonation or man-in-the-middle (MitM) attacks. Mega meets this problem by providing a hierarchical authentication mechanism for contacts and their keys. To avoid any hassle when using multiple types of key...

  8. Aircushion Supported Mega-Floaters

    Van Kessel, J.L.F.

    2010-01-01

    The increase of the global population and expanding coastal mega-cities will necessitate an innovative pursuit of the utilization of the ocean space in which mega-floaters will play an important role in the future. These types of structures are very large floating artificial islands that can be used

  9. Photon detector for MEGA

    During this past August and September, we had beam time at LAMPF for an engineering study of the second prototype cylindrical photon pair spectrometer for MEGA. All of the scintillators in the detector, a total of 40, and 40% of the drift chamber cells were instrumented for this run. The main photon arm activities during the run were to compare event patterns in the chamber to our Monte Carlo generated events, to study the trigger rate and to determine the background rates in the various detector elements. At low beam intensity, the event patterns from the chamber closely resembled those generated from the Monte Carlo. The background rates in the scintillators and the innermost drift chamber layer were close to those anticipated from previous studies. However the background rates in the outer two drift chamber layers were substantially higher than we had expected. This high rate was traced to low energy photons interacting with field and sense wires. The trigger studies during the run have led us to consider alternative strategies including two different first stage triggers and a second stage trigger. The combination of the second stage trigger with either of the two first stage triggers is expected to provide good detection efficiency while keeping the raw trigger rate below that required by the data acquisition system. Detailed discussions of both the background and trigger studies are discussed in this report. Since the run, our work on methods to obtain the z-position in the photon arm drift chambers has continued. Our goal is to obtain the z coordinate to 5 mm FWHM. At this level, the z uncertainty makes a negligible contribution to the overall photon energy resolution and only a small contribution to the angular resolution. We have been studying an option which uses delay lines to provide a direct z determination. The results of our study are discussed in this report

  10. Mega-constellations Issues

    Bastida Virgili, Benjamin; Krag, Holger

    2016-07-01

    increase drastically. In this study, we report the detected issues of such a mega-constellation traffic, and we analyse the response of the space object population to the introduction of a large constellation conforming to the post-mission disposal guideline with differing levels of success and with different disposal orbit options.

  11. Mega cultural events: Does attendance affect residents’ perceptions of a city’s identity?

    Santos, José Freitas; Vareiro, Laurentina; Remoaldo, Paula Cristina Almeida; Ribeiro, J. Cadima

    2015-01-01

    Events are frequently planned with a goal of attracting tourists to a destination. However, studies about the impact of a mega cultural event on residents’ identity are scarce. This research is based on the premise that hosting a mega cultural event can, to a significant degree, change a few of the attributes of the city’s identity as it is perceived by its residents. Specifically, the objective of the study is to identify the attributes of the city and compare the differences found between d...

  12. Five Predatory Mega-Journals: A Review

    Beall, Jeffrey

    2013-01-01

    Mega-journals such as PLOS One are an emerging and successful model of scholarly Open Access publishing. Unfortunately, some new, questionable journals have appeared that are copying the megajournal model. This review covers the five predatory mega-journals British Journal of Science, International Journal of Current Research, International Journal of Science and Advanced Technology (IJSAT), International Journal of Sciences (IJSciences), and World Journal of Science and Technology. Each of t...

  13. Mega-supramolecules for safer, cleaner fuel

    Kornfield, Julie

    Guided by the statistical mechanics of ring-chain equilibrium, we designed and synthesized polymers that self-assemble into ``mega-supramolecules'' (>=5,000 kg/mol) at low concentration (400 kg/mol) and end-association strength is optimal (16-18kT). Hydrocarbon liquid fuels are the world's dominant power source (34% of global energy consumption). Transportation relies heavily on such liquids, presenting the risk of explosive post-impact fires. The collapse of the World Trade Center on September 11, 2001 inspired us to revisit polymers for mist control to mitigate post-impact fuel explosions. Rheological and both light and neutron scattering measurements of long end-functional polymers having polycyclooctadiene backbones and acid or amine end groups verify formation of mega-supramolecules. Post-impact flame propagations experiments show that mega-supramolecules control misting. Turbulent flow measurements show that mega-supramolecules reduce drag like ultra-long covalent polymers. With individual building blocks short enough to avoid hydrodynamic chain scission (400filters without degradation. Mega-supramolecules had no adverse effect on power output, fuel efficiency or emissions in diesel engines. In fact, they gave a 12% reduction in diesel soot. Thus, long end-associative polymers may open the way to fuel additives that reduce pollution and improve transportation safety and security.

  14. Plasmid segregation mechanisms

    Ebersbach, Gitte; Gerdes, Kenn; Charbon, Gitte Ebersbach

    2005-01-01

    Bacterial plasmids encode partitioning (par) loci that ensure ordered plasmid segregation prior to cell division. par loci come in two types: those that encode actin-like ATPases and those that encode deviant Walker-type ATPases. ParM, the actin-like ATPase of plasmid R1, forms dynamic filaments...... that segregate plasmids paired at mid-cell to daughter cells. Like microtubules, ParM filaments exhibit dynamic instability (i.e., catastrophic decay) whose regulation is an important component of the DNA segregation process. The Walker box ParA ATPases are related to MinD and form highly dynamic, oscillating...... filaments that are required for the subcellular movement and positioning of plasmids. The role of the observed ATPase oscillation is not yet understood. However, we propose a simple model that couples plasmid segregation to ParA oscillation. The model is consistent with the observed movement...

  15. SPP1-mediated plasmid transduction.

    Canosi, U; Lüder, G; Trautner, T A

    1982-01-01

    The virulent Bacillus subtilis phage SPP1 transduces plasmid DNA. Plasmid-transducing phages contain only plasmid DNA. Such DNA represents a concatemer of monomeric plasmid molecules with the molecular weight of mature SPP1 DNA. Biological parameters of plasmid transduction are described.

  16. Indian manpower for mega nuclear project

    2003-01-01

    "India is supplying critical scientific manpower and high-tech components needed for building a Large Hadron Collider (LHC) - an accelerator used in particle physics research - a mega scientific project of the European Organisation for Nuclear Research (CERN) worth billions of dollars" (1/2 page).

  17. About urban mega regions : knowns and unknowns

    Yusuf, Shahid

    2007-01-01

    Mega urban regions are not a passing phenomenon. They are likely to persist and to enlarge their economic footprints because they benefit from the advantages of market scale, agglomeration economies, location, and the increasing concentration of talented workers. Metropolitan regions which are polycentric, relatively well managed, and have invested heavily in transport infrastructure are able ...

  18. Construction and Use of Flow Cytometry Optimized Plasmid-Sensor Strains

    Bahl, Martin Iain; Oregaard, Gunnar; Sørensen, Søren Johannes;

    2009-01-01

    Determining the stability of plasmids in bacterial populations is traditionally performed by isolating a large number of clones followed by screening for the presence of plasmids by replica transfer to plasmid-selective agar plates. This is often a laborious task, especially when the intrinsic...... stability of the plasmid is high. The method presented here relies on a phenotypic (green fluorescence protein) marker, which is switched on if the host bacteria loses the residing plasmid. The incorporation of flow cytometry for single-cell detection and discrimination between plasmid-free and plasmid......-harboring cells in a bacterial population facilitates a very high throughput of cells and thus provides excellent sensitivity and statistics toward detecting even very low levels of plasmid instability....

  19. The development of plasmid-free strains of Agrobacterium tumefaciens by using incompatibility with a Rhizobium meliloti plasmid to eliminate pAtC58.

    Hynes, M F; Simon, R; Pühler, A

    1985-03-01

    Agrobacterium tumefaciens strains LBA275 and LBA290 were cured of their cryptic plasmid pAtC58 by the introduction of the Rhizobium meliloti plasmid pRme41a, which is incompatible with pAtC58. pRme41a and pTiC58, the resident Ti plasmid of LBA275, were subsequently eliminated by growth at supraoptimal temperature (40 degrees C). The resulting plasmid-free Agrobacterium strains, UBAPF1 and UBAPF2, have proved extremely useful for the study of Rhizobium plasmids. The loss of the cryptic plasmid pAtC58 has no effect on the tumor-forming ability of the Agrobacterium strains; when the Ti plasmid is present, normal tumors are formed on Kalanchoe daigremontiana. PMID:4001194

  20. Overlap extension PCR cloning: a simple and reliable way to create recombinant plasmids.

    Bryksin, Anton V; Matsumura, Ichiro

    2010-06-01

    Here we describe a straightforward, efficient, and reliable way to clone an insert of choice into a plasmid of choice without restriction endonucleases or T4 DNA ligase. Chimeric primers containing plasmid sequence at the 5' ends and insert sequence at the 3' ends were used to PCR-amplify insertion sequences of various sizes, namely the genes for GFP (gfp), beta-d-glucuronidase (gusA), and beta-galactosidase (lacZ), as well as the entire luxABCDE operon. These inserts were employed as mega-primers in a second PCR with a circular plasmid template. The original plasmid templates were then destroyed in restriction digests with DpnI, and the overlap extension PCR products were used to transform competent Escherichia coli cells. Phusion DNA polymerase was used for the amplification and fusion reactions, so both reactions were easy to monitor and optimize. PMID:20569222

  1. Knowledge Discovery in Mega-Spectra Archives

    Škoda, Petr; Bromová, P.; Lopatovský, L.; Palička, V.; Vážný, J.

    San Francisco: Astronomical Society of the Pacific, 2015, s. 87-90. (ASP Conference Series. 495). ISBN 9781583818749. [Astronomical Data Analysis Software and Systems /24./. Calgary (CA), 05.10.2014-09.10.2014] R&D Projects: GA ČR GA13-08195S Institutional support: RVO:67985815 Keywords : mega-spectra archives * machine learning techniques Subject RIV: BN - Astronomy, Celestial Mechanics, Astrophysics

  2. Vishal Mega-Mart- An Overview

    Shikha Gupta; Preeti Khatri; Kapil Gulati; Santosh Chauhan

    2010-01-01

    In the background of high consumerism and income of the urban consumers, in recent year, a number of companies have expressed their interest towards retail sector outlets. As a result, numbers of shopping malls have started their operations in metro and urban areas. Pantaloon, big bazaar, Vishal Mega Mart, Reliance Fresh are the best known examples of retail sector outlets in India. Retailing is the interface between the producer and the individual consumer buying for personal consumption. Th...

  3. Overlap extension PCR cloning: a simple and reliable way to create recombinant plasmids

    Bryksin, Anton V.; Matsumura, Ichiro

    2010-01-01

    Here we describe a straightforward, efficient, and reliable way to clone an insert of choice into a plasmid of choice without restriction endonucleases or T4 DNA ligase. Chimeric primers containing plasmid sequence at the 5′ ends and insert sequence at the 3′ ends were used to PCR-amplify insertion sequences of various sizes, namely the genes for GFP (gfp), β-D-glucuronidase (gusA), and β-galactosidase (lacZ), as well as the entire luxABCDE operon. These inserts were employed as mega-primers ...

  4. Using Mahalanobis distance to compare genomic signatures between bacterial plasmids and chromosomes

    Suzuki, Haruo; Sota, Masahiro; Brown, Celeste J.; Top, Eva M.

    2008-01-01

    Plasmids are ubiquitous mobile elements that serve as a pool of many host beneficial traits such as antibiotic resistance in bacterial communities. To understand the importance of plasmids in horizontal gene transfer, we need to gain insight into the ‘evolutionary history’ of these plasmids, i.e. the range of hosts in which they have evolved. Since extensive data support the proposal that foreign DNA acquires the host's nucleotide composition during long-term residence, comparison of nucleoti...

  5. Plasmids encoding therapeutic agents

    Keener, William K.

    2007-08-07

    Plasmids encoding anti-HIV and anti-anthrax therapeutic agents are disclosed. Plasmid pWKK-500 encodes a fusion protein containing DP178 as a targeting moiety, the ricin A chain, an HIV protease cleavable linker, and a truncated ricin B chain. N-terminal extensions of the fusion protein include the maltose binding protein and a Factor Xa protease site. C-terminal extensions include a hydrophobic linker, an L domain motif peptide, a KDEL ER retention signal, another Factor Xa protease site, an out-of-frame buforin II coding sequence, the lacZ.alpha. peptide, and a polyhistidine tag. More than twenty derivatives of plasmid pWKK-500 are described. Plasmids pWKK-700 and pWKK-800 are similar to pWKK-500 wherein the DP178-encoding sequence is substituted by RANTES- and SDF-1-encoding sequences, respectively. Plasmid pWKK-900 is similar to pWKK-500 wherein the HIV protease cleavable linker is substituted by a lethal factor (LF) peptide-cleavable linker.

  6. Toxin plasmids of Clostridium perfringens.

    Li, Jihong; Adams, Vicki; Bannam, Trudi L; Miyamoto, Kazuaki; Garcia, Jorge P; Uzal, Francisco A; Rood, Julian I; McClane, Bruce A

    2013-06-01

    In both humans and animals, Clostridium perfringens is an important cause of histotoxic infections and diseases originating in the intestines, such as enteritis and enterotoxemia. The virulence of this Gram-positive, anaerobic bacterium is heavily dependent upon its prolific toxin-producing ability. Many of the ∼16 toxins produced by C. perfringens are encoded by large plasmids that range in size from ∼45 kb to ∼140 kb. These plasmid-encoded toxins are often closely associated with mobile elements. A C. perfringens strain can carry up to three different toxin plasmids, with a single plasmid carrying up to three distinct toxin genes. Molecular Koch's postulate analyses have established the importance of several plasmid-encoded toxins when C. perfringens disease strains cause enteritis or enterotoxemias. Many toxin plasmids are closely related, suggesting a common evolutionary origin. In particular, most toxin plasmids and some antibiotic resistance plasmids of C. perfringens share an ∼35-kb region containing a Tn916-related conjugation locus named tcp (transfer of clostridial plasmids). This tcp locus can mediate highly efficient conjugative transfer of these toxin or resistance plasmids. For example, conjugative transfer of a toxin plasmid from an infecting strain to C. perfringens normal intestinal flora strains may help to amplify and prolong an infection. Therefore, the presence of toxin genes on conjugative plasmids, particularly in association with insertion sequences that may mobilize these toxin genes, likely provides C. perfringens with considerable virulence plasticity and adaptability when it causes diseases originating in the gastrointestinal tract. PMID:23699255

  7. Plasmid-Chromosome Recombination of Irradiated Shuttle Vector DNA in African Green Monkey Kidney Cells.

    Mudgett, John Stuart

    1987-09-01

    An autonomously replicating shuttle vector was used to investigate the enhancement of plasmid-chromosome recombination in mammalian host cells by ultraviolet light and gamma radiation. Sequences homologous to the shuttle vector were stably inserted into the genome of African Green Monkey kidney cells to act as the target substrate for these recombination events. The SV40- and pBR322-derived plasmid DNA was irradiated with various doses of radiation before transfection into the transformed mammalian host cells. The successful homologous transfer of the bacterial ampicillin resistance (amp^{rm r}) gene from the inserted sequences to replace a mutant amp^->=ne on the shuttle vector was identified by plasmid extraction and transformation into E. coli host cells. Ultraviolet light (UV) was found not to induce homologous plasmid-chromosome recombination, while gamma radiation increased the frequency of recombinant plasmids detected. The introduction of specific double -strand breaks in the plasmid or prolonging the time of plasmid residence in the mammalian host cells also enhanced plasmid-chromosome recombination. In contrast, plasmid mutagenesis was found to be increased by plasmid UV irradiation, but not to change with time. Plasmid survival, recombination, and mutagenesis were not affected by treating the mammalian host cells with UV light prior to plasmid transfection. The amp^{rm r} recombinant plasmid molecules analyzed were found to be mostly the result of nonconservative exchanges which appeared to involve both homologous and possibly nonhomologous interactions with the host chromosome. The observation that these recombinant structures were obtained from all of the plasmid alterations investigated suggests a common mechanistic origin for plasmid -chromosome recombination in these mammalian cells.

  8. Likelihood Analysis for Mega Pixel Maps

    Kogut, Alan J.

    1999-01-01

    The derivation of cosmological parameters from astrophysical data sets routinely involves operations counts which scale as O(N(exp 3) where N is the number of data points. Currently planned missions, including MAP and Planck, will generate sky maps with N(sub d) = 10(exp 6) or more pixels. Simple "brute force" analysis, applied to such mega-pixel data, would require years of computing even on the fastest computers. We describe an algorithm which allows estimation of the likelihood function in the direct pixel basis. The algorithm uses a conjugate gradient approach to evaluate X2 and a geometric approximation to evaluate the determinant. Monte Carlo simulations provide a correction to the determinant, yielding an unbiased estimate of the likelihood surface in an arbitrary region surrounding the likelihood peak. The algorithm requires O(N(sub d)(exp 3/2) operations and O(Nd) storage for each likelihood evaluation, and allows for significant parallel computation.

  9. Mega electrorheological phenomena in graphene nanogels

    Dhar, Purbarun; Pattamatta, Arvind; Das, Sarit K

    2016-01-01

    Unprecedentedly massive electrorheology has been reported for dilute graphene nanoflakes based ER fluids that have been engineered as novel, readily synthesizable polymeric gels. Polyethylene glycol based graphene gels have been synthesized and very high ER response, 25,000 percent enhancement in viscosity under influence of electric field, has been observed for low concentration systems 2 wt. percent. The gels overcome several drawbacks innate to ER fluids. The gels exhibit long term stability, high graphene packing ratio which ensures very high ER response and the microstructure of the gels ensure that fibrillation of the graphene nanoflakes under field is undisturbed by thermal fluctuations, further leading to mega ER. The gels exhibit large yield stress handling caliber with yield stress observed as high as 13 kPa at 2 wt. perc. graphene. Detailed investigations on the effects of graphene concentration, electric field strength, imposed shear resistance and transients of electric field actuation on the ER ...

  10. Growth dependence of conjugation explains limited plasmid invasion in biofilms: an individual‐based modelling study

    Merkey, Brian; Lardon, Laurent; Seoane, Jose Miguel;

    2011-01-01

    Plasmid invasion in biofilms is often surprisingly limited in spite of the close contact of cells in a biofilm. We hypothesized that this poor plasmid spread into deeper biofilm layers is caused by a dependence of conjugation on the growth rate (relative to the maximum growth rate) of the donor....... By extending an individual‐based model of microbial growth and interactions to include the dynamics of plasmid carriage and transfer by individual cells, we were able to conduct in silico tests of this and other hypotheses on the dynamics of conjugal plasmid transfer in biofilms. For a generic model plasmid......, we find that invasion of a resident biofilm is indeed limited when plasmid transfer depends on growth, but not so in the absence of growth dependence. Using sensitivity analysis we also find that parameters related to timing (i.e. a lag before the transconjugant can transfer, transfer proficiency...

  11. Permanent resident

    Fisher, John F.

    2016-01-01

    The training of physicians in the past century was based primarily on responsibility and the chain-of-command. Those with the bulk of that responsibility in the fields of pediatrics and internal medicine were residents. Residents trained the medical students and supervised them carefully in caring for patients. Most attending physicians supervised their teams at arm's length, primarily serving as teachers of the finer points of diagnosis and treatment during set periods of the day or week wit...

  12. Persistence Mechanisms of Conjugative Plasmids

    Bahl, Martin Iain; Hansen, Lars H.; Sørensen, Søren Johannes

    2009-01-01

    maintenance in the host cell. These importantly include the ability to self-mobilize in a process termed conjugative transfer, which may occur across species barriers. Other plasmid stabilizing mechanisms include the multimer resolution system, active partitioning, and post-segregational-killing of plasmid......Are plasmids selfish parasitic DNA molecules or an integrated part of the bacterial genome? This chapter reviews the current understanding of the persistence mechanisms of conjugative plasmids harbored by bacterial cells and populations. The diversity and intricacy of mechanisms affecting the...... successful propagation and long-term continued existence of these extra-chromosomal elements is extensive. Apart from the accessory genetic elements that may provide plasmid-harboring cells a selective advantage, special focus is placed on the mechanisms conjugative plasmids employ to ensure their stable...

  13. Mega Construction Projects: using stakeholder management for enhanced sustainable construction

    Eyiah-Botwe, E; Aigbavboa, C; Thwala, WD

    2016-01-01

    The need for mega construction projects development for accelerated socio-economic growth is long overdue for developing countries. Some studies have considered stakeholder management and sustainable construction concepts separately without exploring the simultaneous use for mega projects delivery. This paper reviews the two concepts and suggests a process for enhanced sustainable construction delivery. This study forms part of a broader Ph.D. study on “Development of sustainable ...

  14. In Silico Detection and Typing of Plasmids using PlasmidFinder and Plasmid Multilocus Sequence Typing

    Carattoli, Alessandra; Zankari, Ea; García-Fernández, Aurora; Larsen, Mette Voldby; Lund, Ole; Villa, Laura; Aarestrup, Frank Møller; Hasman, Henrik

    2014-01-01

    genomes of multidrug-resistant Enterobacteriaceae species by the rapid detection of known plasmid types. Replicon sequences from 559 fully sequenced plasmids associated with the family Enterobacteriaceae in the NCBI nucleotide database were collected to build a consensus database for integration into a...... plasmid sequence types (STs) and new alleles and ST variants. In conclusion, testing of the two Web tools using both fully assembled plasmid sequences and WGS-generated draft genomes showed them to be able to detect a broad variety of plasmids that are often associated with antimicrobial resistance in......In the work presented here, we designed and developed two easy-to-use Web tools for in silico detection and characterization of whole-genome sequence (WGS) and whole-plasmid sequence data from members of the family Enterobacteriaceae. These tools will facilitate bacterial typing based on draft...

  15. Permanent resident

    Fisher, John F.

    2016-01-01

    The training of physicians in the past century was based primarily on responsibility and the chain-of-command. Those with the bulk of that responsibility in the fields of pediatrics and internal medicine were residents. Residents trained the medical students and supervised them carefully in caring for patients. Most attending physicians supervised their teams at arm's length, primarily serving as teachers of the finer points of diagnosis and treatment during set periods of the day or week with a perfunctory signature on write-ups or progress notes. Residents endeavored to protect the attending physician from being heavily involved unless they were unsure about a clinical problem. Before contacting the attending physician, a more senior resident would be called. Responsibility was the ultimate teacher. The introduction of diagnosis-related groups by the federal government dramatically changed the health care delivery system, placing greater emphasis on attending physician visibility in the medical record, ultimately resulting in more attending physician involvement in day-to-day care of patients in academic institutions. Without specified content in attending notes, hospital revenues would decline. Although always in charge technically, attending physicians increasingly have assumed the role once dominated by the resident. Using biographical experiences of more than 40 years, the author acknowledges and praises the educational role of responsibility in his own training and laments its declining role in today's students and house staff. PMID:27193992

  16. Mutations in a Partitioning Protein and Altered Chromatin Structure at the Partitioning Locus Prevent Cohesin Recruitment by the Saccharomyces cerevisiae Plasmid and Cause Plasmid Missegregation

    Yang, Xian-Mei; Mehta, Shwetal; Uzri, Dina; Jayaram, Makkuni; Velmurugan, Soundarapandian

    2004-01-01

    The 2μm circle is a highly persistent “selfish” DNA element resident in the Saccharomyces cerevisiae nucleus whose stability approaches that of the chromosomes. The plasmid partitioning system, consisting of two plasmid-encoded proteins, Rep1p and Rep2p, and a cis-acting locus, STB, apparently feeds into the chromosome segregation pathway. The Rep proteins assist the recruitment of the yeast cohesin complex to STB during the S phase, presumably to apportion the replicated plasmid molecules eq...

  17. Conjugative plasmids of Neisseria gonorrhoeae.

    Emilia Pachulec

    Full Text Available Many clinical isolates of the human pathogen Neisseria gonorrhoeae contain conjugative plasmids. The host range of these plasmids is limited to Neisseria species, but presence of a tetracycline (tetM determinant inserted in several of these plasmids is an important cause of the rapid spread of tetracycline resistance. Previously plasmids with different backbones (Dutch and American type backbones and with and without different tetM determinants (Dutch and American type tetM determinants have been identified. Within the isolates tested, all plasmids with American or Dutch type tetM determinants contained a Dutch type plasmid backbone. This demonstrated that tetM determinants should not be used to differentiate between conjugal plasmid backbones. The nucleotide sequences of conjugative plasmids with Dutch type plasmid backbones either not containing the tetM determinant (pEP5233 or containing Dutch (pEP5289 or American (pEP5050 type tetM determinants were determined. Analysis of the backbone sequences showed that they belong to a novel IncP1 subfamily divergent from the IncP1alpha, beta, gamma, delta and epsilon subfamilies. The tetM determinants were inserted in a genetic load region found in all these plasmids. Insertion was accompanied by the insertion of a gene with an unknown function, and rearrangement of a toxin/antitoxin gene cluster. The genetic load region contains two toxin/antitoxins of the Zeta/Epsilon toxin/antitoxin family previously only found in Gram positive organisms and the virulence associated protein D of the VapD/VapX toxin/antitoxin family. Remarkably, presence of VapX of pJD1, a small cryptic neisserial plasmid, in the acceptor strain strongly increased the conjugation efficiency, suggesting that it functions as an antitoxin for the conjugative plasmid. The presence of the toxin and antitoxin on different plasmids might explain why the host range of this IncP1 plasmid is limited to Neisseria species. The isolated plasmids

  18. A plasmid in the archaebacterium Sulfolobus acidocaldarius

    Yeats, Siobhán; McWilliam, Peter; Zillig, Wolfram

    1982-01-01

    A plasmid of mol. wt. ∼9 × 106 has been isolated from the archaebacterium Sulfolobus acidocaldarius strain B12. Plasmid production is induced by u.v. radiation. A copy of the plasmid is probably carried by the chromosome, integrated at a specific site. The entire plasmid, and also restriction fragments of it, has been cloned into Escherichia coli plasmid vectors, and the cleavage sites on the plasmid DNA of three restriction endonucleases have been mapped.

  19. Plasmid acquisition in microgravity

    Juergensmeyer, Margaret A.; Juergensmeyer, Elizabeth A.; Guikema, James A.

    1995-01-01

    In microgravity, bacteria often show an increased resistance to antibiotics. Bacteria can develop resistance to an antibiotic after transformation, the acquisition of DNA, usually in the form of a plasmid containing a gene for resistance to one or more antibiotics. In order to study the capacity of bacteria to become resistant to antibiotics in microgravity, we have modified the standard protocol for transformation of Escherichia coli for use in the NASA-flight-certified hardware package, The Fluid Processing Apparatus (FPA). Here we report on the ability of E. coli to remain competent for long periods of time at temperatures that are readily available on the Space Shuttle, and present some preliminary flight results.

  20. Evolved plasmid-host interactions reduce plasmid interference cost.

    Yano, Hirokazu; Wegrzyn, Katarznya; Loftie-Eaton, Wesley; Johnson, Jenny; Deckert, Gail E; Rogers, Linda M; Konieczny, Igor; Top, Eva M

    2016-09-01

    Antibiotic selection drives adaptation of antibiotic resistance plasmids to new bacterial hosts, but the molecular mechanisms are still poorly understood. We previously showed that a broad-host-range plasmid was poorly maintained in Shewanella oneidensis, but rapidly adapted through mutations in the replication initiation gene trfA1. Here we examined if these mutations reduced the fitness cost of TrfA1, and whether this was due to changes in interaction with the host's DNA helicase DnaB. The strains expressing evolved TrfA1 variants showed a higher growth rate than those expressing ancestral TrfA1. The evolved TrfA1 variants showed a lower affinity to the helicase than ancestral TrfA1 and were no longer able to activate the helicase at the oriV without host DnaA. Moreover, persistence of the ancestral plasmid was increased upon overexpression of DnaB. Finally, the evolved TrfA1 variants generated higher plasmid copy numbers than ancestral TrfA1. The findings suggest that ancestral plasmid instability can at least partly be explained by titration of DnaB by TrfA1. Thus under antibiotic selection resistance plasmids can adapt to a novel bacterial host through partial loss of function mutations that simultaneously increase plasmid copy number and decrease unfavorably high affinity to one of the hosts' essential proteins. PMID:27121483

  1. Measuring Polycentricity of Mega-City Regions in China Based on the Intercity Migration Flows

    Mu, Xiaoyan; Yeh, Anthony G. O.

    2016-06-01

    This paper uses the intercity migration flows to examine relations between Chinese cities, identify the important mega-city regions and measure each region's polycentricity from an interaction perspective. Data set contains the long-term residential migration trajectories of three million Sina weibo users across 345 cities. Cities with close connectivity deployed around one or several mega cities are identified as mega-city regions. Features of the mega-city regions are characterized by the strength of migration flows, density of connections, and regional migration patterns. The results show that the disparities exist in different mega-city regions; most mega-city regions are lack of polycentricity.

  2. Defining a quantitative framework for evaluation and optimisation of the environmental impacts of mega-event projects.

    Parkes, Olga; Lettieri, Paola; Bogle, I David L

    2016-02-01

    This paper presents a novel quantitative methodology for the evaluation and optimisation of the environmental impacts of the whole life cycle of a mega-event project: construction and staging the event and post-event site redevelopment and operation. Within the proposed framework, a mathematical model has been developed that takes into account greenhouse gas (GHG) emissions resulting from use of transportation fuel, energy, water and construction materials used at all stages of the mega-event project. The model is applied to a case study - the London Olympic Park. Three potential post-event site design scenarios of the Park have been developed: Business as Usual (BAU), Commercial World (CW) and High Rise High Density (HRHD). A quantitative summary of results demonstrates that the highest GHG emissions associated with the actual event are almost negligible compared to those associated with the legacy phase. The highest share of emissions in the legacy phase is attributed to embodied emissions from construction materials (almost 50% for the BAU and HRHD scenarios) and emissions resulting from the transportation of residents, visitors and employees to/from the site (almost 60% for the CW scenario). The BAU scenario is the one with the lowest GHG emissions compared to the other scenarios. The results also demonstrate how post-event site design scenarios can be optimised to minimise the GHG emissions. The overall outcomes illustrate how the proposed framework can be used to support decision making process for mega-event projects planning. PMID:26686076

  3. The Mega-Terrorism - The Challenge of the Third Millennium

    Daniel Andrișan

    2011-06-01

    Full Text Available The aim of this article is to describe the "new face" of terrorism in the twenty-first centuryindicating the transition to mass destruction weapons (nuclear terrorism, chemical, biological, toethnic-religious terrorism, the occurrence of massive gray area phenomena of terrorism and how thisphenomenon became a strategic weapon. Mega-terrorism has been existing, at a conceptual level,ever since the 70’s, when the experts of this phenomenon tried to find a semantic cover for thesituations that certain organizations, groups or terrorist or extremist sections would get someWeapons of Mass Destruction Systems. Actually, the 11th of September tragedy proves that megaterrorismrepresents the premeditated destruction, the lack of negotiations, of a tactical goal that wasto be accomplished by means of a threat. The mega-terrorism actors do not intend to get themselvesknown to the public, do not require anything, do not see the threat as means or device to reach theirpurpose. The 21 st century mega-terrorism simple logic is materialized in the systemic destroying ofthe enemy, no longer representing a political power or a government, but an entire ‘disobeying’,‘corrupted’, ‘unfaithful’ population indifferent to the mega-terrorist actors’ desires. New dimensionagainst mega-terrorism determines not only a rethinking of how to combat it, but also the realizationof a conceptual legal framework and measures established to materialize the political decision.

  4. A new analysis of the MEGA M31 microlensing events

    Ingrosso, G; De Paolis, F; Jetzer, P; Nucita, A A; Scarpetta, G; Strafella, F; Jetzer, Ph.

    2006-01-01

    We discuss the results of the MEGA microlensing campaign towards M31. Our analysis is based on an analytical evaluation of the microlensing rate, taking into account the observational efficiency as given by the MEGA collaboration. In particular, we study the spatial and time duration distributions of the microlensing events for several mass distribution models of the M31 bulge. We find that only for extreme models of the M31 luminous components it is possible to reconcile the total observed MEGA events with the expected self-lensing contribution. Nevertheless, the expected spatial distribution of self-lensing events is more concentrated and hardly in agreement with the observed distribution. We find it thus difficult to explain all events as being due to self-lensing alone. On the other hand, the small number of events does not yet allow to draw firm conclusions on the halo dark matter fraction in form of MACHOs.

  5. Mega-masers, Dark Energy and the Hubble Constant

    Powerful water maser emission (water mega-masers) can be found in accretion disks in the nuclei of some galaxies. Besides providing a measure of the mass at the nucleus, such mega-masers can be used to determine the distance to the host galaxy, based on a kinematic model. We will explain the importance of determining the Hubble Constant to high accuracy for constraining the equation of state of Dark Energy and describe the Mega-maser Cosmology Project that has the goal of determining the Hubble Constant to better than 3%. Time permitting, we will also present the scientific capabilities of the current and future NRAO facilities: ALMA, EVLA, VLBA and GBT, for addressing key astrophysical problems

  6. Mega-masers, Dark Energy and the Hubble Constant

    Lo, Fred K. Y.

    2007-10-15

    Powerful water maser emission (water mega-masers) can be found in accretion disks in the nuclei of some galaxies. Besides providing a measure of the mass at the nucleus, such mega-masers can be used to determine the distance to the host galaxy, based on a kinematic model. We will explain the importance of determining the Hubble Constant to high accuracy for constraining the equation of state of Dark Energy and describe the Mega-maser Cosmology Project that has the goal of determining the Hubble Constant to better than 3%. Time permitting, we will also present the scientific capabilities of the current and future NRAO facilities: ALMA, EVLA, VLBA and GBT, for addressing key astrophysical problems

  7. Characterization of the particulate air pollution in contrasted mega cities

    This work aims at characterizing the physics and the chemistry that govern particulate air pollution in two mega cities (Paris and Cairo) for which the size distribution and the chemical composition of airborne particles were poorly documented. Seasonal variations of the main aerosol sources and transformation processes are investigated in these two urban centres, with a particular attention to semi-volatile material and secondary organic aerosols. Short-term health effects of Paris size-segregated aerosols, as well as particulate pollution during the Cairo 'Black Cloud' season, are also emphasized here. Finally, the comparison of results obtained for the two mega cities and for another one (Beijing) allows investigating main factors responsible for particulate air pollution in urban centres with contrasted climatic conditions and development levels. Notably, this work also allows the build-up of an experimental dataset which is now available for the modelling of urban air quality and of environmental impacts of mega city air pollution. (author)

  8. The Java-Sumatra Aerial Mega-Tramway

    Bolonkin, Alexander; Cathcart, Richard

    2007-01-01

    A mega-tramway based on the Indonesian islands of Sumatra and Java is proposed to span Sunda Strait. The Java-Sumatra Aerial Mega-Tramway (JSAMT) will be self-elevating and will regularly and cheaply launch passengers and cargoes via two conveyor belt-like facilities using standard winged shipping containers like those currently used by international trucking and ocean shipping industries that are volplaned across the Sunda Strait. The JSAMT will be a self-sustaining toll facility free of any...

  9. Global Analysis and Structural Performance of the Tubed Mega Frame

    ZHANG, Han

    2014-01-01

    The Tubed Mega Frame is a new structure concept for high-rise buildings which is developed by Tyréns. In order to study the structural performance as well as the efficiency of this new concept, a global analysis of the Tubed Mega Frame structure is performed using finite element analysis software ETABS. Besides, the lateral loads that should be applied on the structure according to different codes are also studied. From the design code study for wind loads and seismic design response spectrum...

  10. Plasmid-mediated transformation in Bacillus megaterium.

    Brown, B. J.; Carlton, B C

    1980-01-01

    A transformation system was developed for Bacillus megaterium by using antibiotic resistance plasmid deoxyribonucleic acid molecules derived from Staphylococcus aureus and Bacillus cereus. Lysozyme-generated protoplasts of B. megaterium allowed uptake of plasmid deoxyribonucleic acid in the presence of polyethylene glycol. Transformants expressed the antibiotic resistance determinants present on the plasmid deoxyribonucleic acid, and reisolated plasmid deoxyribonucleic acid yielded restrictio...

  11. Strategies simulation model for the gas business chain MEGAS

    MEGAS is a simulation model representing fundamental parameters of Corpoven's natural gas and NGL production, handling, processing, transportation and distribution systems in the Venezuelan mid-east, as well as its financial implications. Various strategies regarding development, prices, costs, new business opportunities, production scenarios, demand and energy policies can be evaluated through this model in order to determine, after analyzing the economics results, a set of strategies to follow in the mid and long term. MEGAS could be also used to make risk analysis studies, considering that probabilistic parameters and variables like gas quality, production, demand, plant shutdowns and others are to be represented by their distinctive function. It is possible to set up a probabilistic function for each economic indicator or operating variables with an appropriate experiment design. MEGAS is based on a dynamic simulation language, which facilitates both the real system components representation and the main variables statistical data accumulation. It also allows graphical representation of results and the simulation animation. Demand and price forecasts, new projects, costs and capabilities of the systems are some of the model input data. MEGAS should be considered as a tool that eases the strategic planning of the business, making it possible for the Corporation to foresee changes, both in the domestic and international market, to predict how these changes could affect its business affairs and to visualize different return scenarios

  12. The partitioning and copy number control systems of the selfish yeast plasmid: an optimized molecular design for stable persistence in host cells

    Yen-Ting-Liu,; Sau, Saumitra; Ma, Chien-Hui; Kachroo, Aashiq H.; Rowley, Paul A.; Chang, Keng-Ming; Fan, Hsiu-Fang; Jayaram, Makkuni

    2014-01-01

    The multi-copy 2 micron plasmid of Saccharomyces cerevisiae, a resident of the nucleus, is remarkable for its high chromosome-like stability. The plasmid does not appear to contribute to the fitness of the host, nor does it impose a significant metabolic burden on the host at its steady state copy number. The plasmid may be viewed as a highly optimized selfish DNA element whose genome design is devoted entirely towards efficient replication, equal segregation and copy number maintenance. A pa...

  13. Effects of genes exerting growth inhibition and plasmid stability on plasmid maintenance.

    Boe, L; Gerdes, K; Molin, S

    1987-01-01

    Plasmid stabilization mediated by the parA+ and parB+ genes of the R1 plasmid and the ccd+ and sop+ genes of the F plasmid was tested on a mini-R1 plasmid and a pBR322 plasmid derivative. The mini-R1 plasmid is thought to be unstably inherited owing to a low copy number and to random segregation of the plasmid at cell division, whereas cells harboring the pBR322 derivative used in this work are lost through competition with plasmid-free cells, mainly as a result of the shorter generation time...

  14. Repair in E. coli of transforming plasmid DNA damaged by psoralen plus near-ultraviolet irradiation

    Treatment of DNA with psoralen plus near-ultraviolet irradiation gives rise to both monoadducts and cross-links. The authors have examined the repair of plasmid NTP16 DNA treated in this way in vitro and then used to transform E. coli. Monoadducts are found to be potentially lethal, and can be repaired by uvr-dependent and recA-dependent pathways. The presence of a related resident plasmid in the transformed cells can enhance the survival of the incoming damaged NTP16 DNA. This effect is not recA-dependent. Removal of unbound psoralen from the plasmid DNA and exposure to further NUV is known to increase the ratio of cross-links to monoadducts, and the authors demonstrate that such cross-linked plasmid DNA is not readily repaired following transformation. However in the presence of homologous DNA there is evidence for the repair, and hence uptake by the cell, of cross-linked DNA. (Auth.)

  15. Genomic and Functional Characterization of qnr-Encoding Plasmids from Municipal Wastewater Biosolid Klebsiella pneumoniae Isolates

    Kaplan, Ella; Sela, Noa; Doron-Faigenboim, Adi; Navon-Venezia, Shiri; Jurkevitch, Edouard; Cytryn, Eddie

    2015-01-01

    -like plasmids concomitant to phylogenetic analysis of housekeeping genes from host Klebsiella strains, revealed that these plasmids are limited to a predominantly human-associated sub-clade of Klebsiella, suggesting that their host range is very narrow. Conversely, the pGNB2-like plasmids had a much broader host range and appeared to be associated with Klebsiella residing in natural environments. This study suggests that: (A) qnrB-harboring multidrug-resistant pKPN3-like plasmids can endure the rigorous wastewater treatment process and may therefore be disseminated to downstream environments; and (B) that small qnrS-harboring pGNB2-like plasmids are ubiquitous in wastewater treatment facilities and are most likely environmental in origin. PMID:26696974

  16. Management of infected custom mega prosthesis by Ilizarov method

    Karthik Gudaru

    2015-12-01

    Full Text Available A 26 year old male patient with an aggressive giant cell tumour of the left distal femur was managed by wide excision and insertion of custom mega prosthesis. Patient developed deep infection which did not subside with multiple debridements and antibiotics. Hence the custom mega prosthesis was removed and the bone gap was managed by Ilizarov method. Ilizarov fixation of the left femur and tibia was done. Femoral and tibial corticotomy was done and the bone segments were transported towards the knee to finally achieve a knee arthrodesis. The case illustrates the method and difficulties encountered during the treatment. [Int J Res Med Sci 2015; 3(12.000: 3874-3878

  17. Astrocyte mega-domain hypothesis of the autistic savantism.

    Mitterauer, Bernhard J

    2013-01-01

    Individuals with autism who show high abilities are called savants. Whereas in their brains a disconnection in and between neural networks has been identified, savantism is yet poorly understood. Focusing on astrocyte domain organization, it is hypothesized that local astrocyte mega-organizations may be responsible for exerting high capabilities in brains of autistic savants. Astrocytes, the dominant glial cell type, modulate synaptic information transmission. Each astrocyte is organized in non-overlapping domains. Formally, each astrocyte contacting n-neurons with m-synapses via its processes generates dynamic domains of synaptic interactions based on qualitative computation criteria, and hereby it structures neuronal information processing. If the number of processes is genetically significantly increased, these astrocytes operate in a mega-domain with a higher complexitiy of computation. From this model savant abilities are deduced. PMID:23098371

  18. Antiparallel plasmid-plasmid pairing may control P1 plasmid replication.

    Abeles, A L; Austin, S J

    1991-01-01

    The copy number of the P1 plasmid replicon is stringently controlled, giving only one or two copies per newborn cell. Control is achieved by the action of the copy-control locus incA, which contains nine repeats of the 19-basepair binding site for the plasmid-encoded initiator protein RepA. A set of five similar repeats are present in the replication origin where RepA acts to trigger initiation. Using an in vitro replication system consisting of an Escherichia coli extract, the P1 origin as a...

  19. Plasmid profiling of bacterial isolates from confined environments

    van Houdt, Rob; Provoost, Ann; Coninx, Ilse; Leys, Natalie; Mergeay, Max

    Plasmid profiling of bacterial isolates from confined environments R. Van Houdt, I. Coninx, A. Provoost, N. Leys, and M. Mergeay Expertise group for Molecular and Cellular Biology, Institute for Environment, Health and Safety, Belgian Nuclear Research Centre (SCK•CEN), Boeretang 200, B-2400 Mol, Belgium. Human exploration of extreme and isolated hostile environments such as space requires special confined small volume habitats to protect and house the crew. However, human confinement in such small volume habitats has restrictions on waste disposal and personal hygiene and inevitably generates a particular community of microorganisms within the habitat. These microorganisms are mainly originating from the crew (skin, mucous membranes, upper respiratory tract, mouth, and gastrointestinal tract) but also include the residing environmental microorganisms. Earth-based confined habitats such as the Antarctic Research Station Concordia are used as test beds for long-duration spaceflights to study the physiologic and psychological adaptation to isolated environments. The dynamics of the environmental microbial population in such a test bed could render additional insights in assessing the potential health risks in long-duration space missions. Not only total bacterial contamination levels are important, but it is essential to identify also the predominant microbial taxa and their mobile genetic elements (MGE). These MGEs could be exchanged between bacteria by horizontal gene transfer and may alter the pathogenic potential since they often carry antibiotic resistance or more in general adaptation-enhancing traits. In this study several bacterial strains isolated in the Concordia research station were examined for their plasmid content. An optimized protocol for extraction of large plasmids showed the present of at least one plasmid in 50% of the strains. For all strains the minimal inhibitory concentration of a range of antibiotics was determined indicating resistance to

  20. A plasmid in Legionella pneumophila.

    Knudson, G B; Mikesell, P

    1980-01-01

    Sixteen strains from the six serogroups of Legionella pneumophila were examined for the presence of extrachromosomal genetic elements by a modified cleared lysate procedure, dye-buoyant centrifugation, and agarose gel electrophoresis. Two strains, Atlanta-1 and Atlanta-2 from serogroup II, each contained a plasmid of cryptic function with a molecular weight of ca. 30 megadaltons.

  1. The Mega-Terrorism - The Challenge of the Third Millennium

    Daniel Andrișan

    2011-01-01

    The aim of this article is to describe the "new face" of terrorism in the twenty-first centuryindicating the transition to mass destruction weapons (nuclear terrorism, chemical, biological), toethnic-religious terrorism, the occurrence of massive gray area phenomena of terrorism and how thisphenomenon became a strategic weapon. Mega-terrorism has been existing, at a conceptual level,ever since the 70’s, when the experts of this phenomenon tried to find a semantic cover for thesituations that ...

  2. ROLE OF MEGA SPORT EVENTS ON BRANDING DESTINATIONS

    Abolfazl Farahani; Habib Honari; Hasan Asadi; Abdul Hamid Zeitoonly; Mohammad Pour Ranjbar

    2011-01-01

    Sporting events to help make creating beautiful scenery and unique areas to tourism. Thepurpose of this study was investigated the role of holding sporting events on destinationsBranding with hosted of mega sports event (Traditional Sport ;hors riding) on branding sportstourism destinations in Golestan province. Data from a questionnaire (Cronbach's alpha = 91%)and interviews were collected. Statistical population of this study was 217 patients (53 expert(cultural heritage; physical education...

  3. Some Aspects of Mega-Floating Airport Design and Production

    Hadžić, Neven; Tomić, Marko; Vladimir, Nikola; Senjanović, Ivo

    2016-01-01

    Mega-Floating Airports (MFA) are unique and complex offshore transport system components that emerged as a consequence of tremendous land price increase in the vicinity of very large coastal cities. An overview of MFAs design and production aspects is presented within this paper including design concept, model tests and full scale measurement, air transport analysis, infrastructure, main particulars and structure, wave breaker, hydroelastic analysis due to wave load and airplane moving ...

  4. Assessment of MEGA BORG impacts on sea turtles

    Studies were conducted to assess the impacts of the MEGA BORG oil spill on sea turtles in the path of the oil plume. Aerial surveys were performed to determine the presence of turtles and provide a gross visual assessment of potential impacts. Although extensive efforts were made to capture sea turtles around oil and gas platforms only one loggerhead sea turtle, Caretta caretta, was captured. Neither external visual inspection nor laboratory fecal analysis showed evidence of petroleum contamination

  5. TICKET UNDERPRICING AND PUBLIC SUPPORT FOR MEGA-EVENTS

    Eichhorn, Christoph; Sahm, Marco

    2010-01-01

    The organizer of a mega-event often owns a monopoly for several products like tickets and sponsorship contracts. While ticket prices below the maximum sellout price reduce gate revenues, they increase the degree to which the event is accepted by the population of the host region. Since public support usually serves as an important input factor for sponsors, it increases revenues in the market for sponsorship contracts. Hence, ticket underpricing may well maximize overall profits of the organi...

  6. Mega borg oil spill: Fate and effect studies

    The Mega Borg, a Norwegian tanker, released an estimated 5.1 million gallons (gal) of Palanca Angola crude oil into the Gulf of Mexico during a lightering accident and subsequent fire. The collection of reports was designed to provide a comprehensive overview of the spill chronology, the fate of the oil released, and subsequent studies that were conducted to assess the impacts of the oil spill on the environment and its biota

  7. Bacterial Plasmids in Antarctic Natural Microbial Assemblages

    Kobori, Hiromi; Sullivan, Cornelius W.; Shizuya, Hiroaki

    1984-01-01

    Samples of psychrophilic and psychrotrophic bacteria were collected from sea ice, seawater, sediments, and benthic or ice-associated animals in McMurdo Sound, Antarctica. A total of 155 strains were isolated and tested for the presence of plasmids by DNA agarose gel electrophoresis. Thirty-one percent of the isolates carried at least one kind of plasmid. Bacterial isolates taken from sediments showed the highest plasmid incidence (42%), and isolates from seawater showed the lowest plasmid inc...

  8. Plasmid maintenance functions encoded on Dictyostelium discoideum nuclear plasmid Ddp1.

    Hughes, J E; H. Kiyosawa; Welker, D L

    1994-01-01

    All of the plasmid-carried genes expressed during vegetative growth are essential for long-term maintenance of plasmid Ddp1 in the nucleus of Dictyostelium discoideum. Deletion of Ddp1 genes expressed only during development had no detectable effect on plasmid maintenance. Deletion of vegetatively expressed genes, either singly or in pairs, resulted in (i) a rapid loss of plasmid from cells grown in the absence of selection for plasmid retention, (ii) variation in the proportion of monomer to...

  9. IncP-1β Plasmid pGNB1 Isolated from a Bacterial Community from a Wastewater Treatment Plant Mediates Decolorization of Triphenylmethane Dyes▿

    Schlüter, Andreas; Krahn, Irene; Kollin, Florian; Bönemann, Gabriele; Stiens, Michael; Szczepanowski, Rafael; Schneiker, Susanne; Pühler, Alfred

    2007-01-01

    Plasmid pGNB1 was isolated from bacteria residing in the activated sludge compartment of a wastewater treatment plant by using a transformation-based approach. This 60-kb plasmid confers resistance to the triphenylmethane dye crystal violet and enables its host bacterium to decolorize crystal violet. Partial sequencing of pGNB1 revealed that its backbone is very similar to that of previously sequenced IncP-1β plasmids. The two accessory regions of the plasmid, one located downstream of the re...

  10. Characterization of plasmids in Erwinia stewartii.

    Coplin, D. L.; Rowan, R G; Chisholm, D A; Whitmoyer, R E

    1981-01-01

    Plasmids in 39 strains of Erwinia stewartii were examined by agarose gel electrophoresis. Most virulent strains had from 11 to 13 plasmids ranging in molecular mass from 2.8 to 210 megadaltons and contained plasmids of 210, 70, 49, 43, 29.5, 16.8, 8.8, and 2.8 megadaltons. Plasmids in strains SW2 and SS104 were characterized by both electron microscopy and agarose gel electrophoresis and may be useful as convenient references for sizing plasmids by electrophoresis. Specific size classes of pl...

  11. Residency Allocation Database

    Department of Veterans Affairs — The Residency Allocation Database is used to determine allocation of funds for residency programs offered by Veterans Affairs Medical Centers (VAMCs). Information...

  12. Mechanism Analysis and Parameter Optimization of Mega-Sub-Isolation System

    Xiangxiu Li; Ping Tan; Xiaojun Li; Aiwen Liu

    2016-01-01

    The equation of motion of mega-sub-isolation system is established. The working mechanism of the mega-sub-isolation system is obtained by systematically investigating its dynamic characteristics corresponding to various structural parameters. Considering the number and location of the isolated substructures, a procedure to optimally design the isolator parameters of the mega-sub-isolation system is put forward based on the genetic algorithm with base shear as the optimization objective. The i...

  13. Social and philosophical conceptualization mega society as the body in the context of global synergetic methodology

    Мельник, Вікторія Володимирівна

    2014-01-01

    This paper analyzes how global body; theoretical and conceptual framework of the society as a single mega social, cultural and global organism; concepts and categories of the mega social context in synergetic methodology as attractors, repeler " cultural circle" , the development of culture in the context mega society; Cultural Dialogue in the global development community. It is proved that cultural interaction seems important structure- factor that binds the human community and discovers ide...

  14. Virulence Plasmids of Spore-Forming Bacteria.

    Adams, Vicki; Li, Jihong; Wisniewski, Jessica A; Uzal, Francisco A; Moore, Robert J; McClane, Bruce A; Rood, Julian I

    2014-12-01

    Plasmid-encoded virulence factors are important in the pathogenesis of diseases caused by spore-forming bacteria. Unlike many other bacteria, the most common virulence factors encoded by plasmids in Clostridium and Bacillus species are protein toxins. Clostridium perfringens causes several histotoxic and enterotoxin diseases in both humans and animals and produces a broad range of toxins, including many pore-forming toxins such as C. perfringens enterotoxin, epsilon-toxin, beta-toxin, and NetB. Genetic studies have led to the determination of the role of these toxins in disease pathogenesis. The genes for these toxins are generally carried on large conjugative plasmids that have common core replication, maintenance, and conjugation regions. There is considerable functional information available about the unique tcp conjugation locus carried by these plasmids, but less is known about plasmid maintenance. The latter is intriguing because many C. perfringens isolates stably maintain up to four different, but closely related, toxin plasmids. Toxin genes may also be plasmid-encoded in the neurotoxic clostridia. The tetanus toxin gene is located on a plasmid in Clostridium tetani, but the botulinum toxin genes may be chromosomal, plasmid-determined, or located on bacteriophages in Clostridium botulinum. In Bacillus anthracis it is well established that virulence is plasmid determined, with anthrax toxin genes located on pXO1 and capsule genes on a separate plasmid, pXO2. Orthologs of these plasmids are also found in other members of the Bacillus cereus group such as B. cereus and Bacillus thuringiensis. In B. thuringiensis these plasmids may carry genes encoding one or more insecticidal toxins. PMID:26104459

  15. Novel archaeal plasmid pAH1 and its interactions with the lipothrixvirus AFV1

    Basta, Tamara; Smyth, John; Forterre, Patrick; Prangishvili, David; Peng, Xu

    2009-01-01

    At present very little is known about interactions between extrachromosomal genetic elements in Archaea. Here we describe an Acidianus strain which carries naturally a novel 28 kb conjugative plasmid-like element, pAH1, and also serves as a laboratory host for lipothrixvirus AFV1. In an attempt t...... observed in bacteria where relevant bacteriophages either are dependent on a conjugative plasmid for successful infection or are excluded by a resident plasmid.......At present very little is known about interactions between extrachromosomal genetic elements in Archaea. Here we describe an Acidianus strain which carries naturally a novel 28 kb conjugative plasmid-like element, pAH1, and also serves as a laboratory host for lipothrixvirus AFV1. In an attempt to...... establish a system for studying plasmid-virus interactions we characterized the genome of pAH1 which closely resembles those of the Sulfolobus conjugative plasmids pARN3 and pARN4. pAH1 integrates site specifically into, and excises from, the host chromosome indicating a dynamic interaction with the latter...

  16. How to make mega-cities energy efficient?

    Aoki, Hitoshi; Aoki, Yoshitaka

    2010-09-15

    Tokyo is a Megalopolis with 40 million+ population. It has an energy efficient profile due to its uniqueness as extensive mass rail transit networks, high density and compact urban formation, compact space oriented life style. The other feature is extensive use of electric heat pumps (EHP), which entails low carbon city profile. Further possibility is prepared with water thermal energy utilization, which is widely available also through EHP particularly in Tokyo central districts, which could make Tokyo one of the lowest carbon cities in the world. Emerging mega-cities are expected to learn from Tokyo's success and not from western conventional models.

  17. ColE1-plasmid production in Escherichia coli: Mathematical Simulation and Experimental Validation

    Inga eFreudenau

    2015-09-01

    Full Text Available Plasmids have become very important as pharmaceutical gene vectors in the fields of gene therapy and genetic vaccination in the last years. In this study, we present a dynamic model to simulate the ColE1-like plasmid replication control, once for a DH5α-strain carrying a low copy plasmid (DH5α-pSUP 201-3 and once for a DH5α-strain carrying a high copy plasmid (DH5α-pCMV-lacZ by using ordinary differential equations (ODE and the MATLAB software. The model includes the plasmid replication control by two regulatory RNA molecules (RNAI and RNAII as well as the replication control by uncharged tRNA molecules. To validate the model, experimental data like RNAI- and RNAII concentration, plasmid copy number (PCN, and growth rate for three different time points in the exponential phase were determined. Depending on the sampled time point, the measured RNAI and RNAII concentrations for DH5α-pSUP 201-3 reside between 6 ±0.7 to 34 ±7 RNAI molecules per cell and 0.44 ±0.1 to 3 ±0.9 RNAII molecules per cell. The determined plasmid copy numbers (PCN averaged between 46 ±26 to 48 ±30 plasmids per cell. The experimentally determined data for DH5α-pCMV-lacZ reside between 345 ±203 to 1086 ±298 RNAI molecules per cell and 22 ±2 to 75 ±10 RNAII molecules per cell with an averaged PCN of 1514 ±1301 to 5806 ±4828 depending on the measured time point. As the model was shown to be consistent with the experimentally determined data, measured at three different time points within the growth of the same strain, we performed predictive simulations concerning the effect of uncharged tRNA molecules on the ColE1-like plasmid replication control. The hypothesis is that these tRNA molecules would have an enhancing effect on the plasmid production. The in silico analysis predicts that uncharged tRNA molecules would indeed increase the pDNA production.

  18. Plasmid and chromosome partitioning: surprises from phylogeny

    Gerdes, Kenn; Møller-Jensen, Jakob; Bugge Jensen, Rasmus

    2000-01-01

    Plasmids encode partitioning genes (par) that are required for faithful plasmid segregation at cell division. Initially, par loci were identified on plasmids, but more recently they were also found on bacterial chromosomes. We present here a phylogenetic analysis of par loci from plasmids...... and chromosomes from prokaryotic organisms. All known plasmid-encoded par loci specify three components: a cis-acting centromere-like site and two trans-acting proteins that form a nucleoprotein complex at the centromere (i.e. the partition complex). The proteins are encoded by two genes in an operon...... that is autoregulated by the par-encoded proteins. In all cases, the upstream gene encodes an ATPase that is essential for partitioning. Recent cytological analyses indicate that the ATPases function as adaptors between a host-encoded component and the partition complex and thereby tether plasmids and chromosomal...

  19. Homemade Site Directed Mutagenesis of Whole Plasmids

    Laible, Mark; Boonrod, Kajohn

    2009-01-01

    Site directed mutagenesis of whole plasmids is a simple way to create slightly different variations of an original plasmid. With this method the cloned target gene can be altered by substitution, deletion or insertion of a few bases directly into a plasmid. It works by simply amplifying the whole plasmid, in a non PCR-based thermocycling reaction. During the reaction mutagenic primers, carrying the desired mutation, are integrated into the newly synthesized plasmid. In this video tutorial we demonstrate an easy and cost effective way to introduce base substitutions into a plasmid. The protocol works with standard reagents and is independent from commercial kits, which often are very expensive. Applying this protocol can reduce the total cost of a reaction to an eighth of what it costs using some of the commercial kits. In this video we also comment on critical steps during the process and give detailed instructions on how to design the mutagenic primers. PMID:19488024

  20. Local meanings of a sport mega-event's legacies : Stories from a South African urban neighbourhood

    Waardenburg, Maikel; van den Bergh, Marjolein; van Eekeren, Frank

    2015-01-01

    Studies on sport mega-events and their legacies often seem only loosely connected to local experiences. Stories on sport mega-event legacy appear as a setting-the-scene or function as a reference to illustrate specific types of legacy. However, stories themselves are never the primary focus in these

  1. Uptake of plasmid deoxyribonucleic acid by Haemophilus.

    Gromkova, R; Goodgal, S

    1981-01-01

    The uptake of circular and linear plasmid RSF0885 deoxyribonucleic acids, (DNAs) obtained from Haemophilus parainfluenzae 14, in both homologous and heterologous recipients was studied and compared with that of chromosomal DNA. High concentrations of divalent cations stimulated the uptake of either circular or linear plasmid DNA in H. parainfluenzae 14 competent cells but did not affect the uptake of chromosomal DNA. The biological activity of linear plasmid DNA was similar to that of circula...

  2. Development of sewage system for mega-city Karachi

    The growth of mega-cities in recent times has brought to the fore a large number of issues with respect environmental conditions such as development of slum areas, poor localities, solid waste disposal, management of sewage system, lack of open spaces and health facilities. One of the major issues effecting environment proper disposal of sewage and development of sewage system including recycling of sewage water and its use. The heavy pressure of population calls for adequate provision of sewage disposal in the new areas and remodeling of systems in the old areas of the cities to ensure a pollution-free environment. Recycling of sewage and prevention of sewage discharge into natural water channels and the sea has to be undertaken on urgent bases. The paper seeks to outline the present status of sewage system of the city of Karachi, the problems involved and the proposals for management and development of the system compatible with the need and requirement of mega-city population. (author)

  3. MegaPipe astrometry for the New Horizons spacecraft

    Gwyn, Stephen D J

    2014-01-01

    The New Horizons spacecraft, launched by NASA in 2006, will arrive in the Pluto-Charon system on July 14, 2015. There, it will spend a few hours imaging Pluto and its moons. It will then have a small amount of reserve propellant which will be used to direct the probe on to a second, yet to be discovered object in the Kuiper Belt. Data from the MegaPrime camera on CFHT was used to build a precise, high density astrometric reference frame for both the final approach into the Pluto system and the search for the secondary target. Pluto currently lies in the galactic plane. This is a hindrance in that there are potential problems with confusion. However, it is also a benefit, since it allows the use of the UCAC4 astrometric reference catalog, which is normally too sparse for use with MegaCam images. The astrometric accuracy of the final catalogs, as measured by the residuals, is 0.02 arcseconds.

  4. Mega Construction Projects: using stakeholder management for enhanced sustainable construction

    Eyiah-Botwe, E

    2016-06-01

    Full Text Available The need for mega construction projects development for accelerated socio-economic growth is long overdue for developing countries. Some studies have considered stakeholder management and sustainable construction concepts separately without exploring the simultaneous use for mega projects delivery. This paper reviews the two concepts and suggests a process for enhanced sustainable construction delivery. This study forms part of a broader Ph.D. study on “Development of sustainable stakeholder management framework for construction projects” aimed at improving stakeholder management process and project delivery. A qualitative research method approach was adopted. An extensive literature review on stakeholder management and sustainable construction was conducted using filtering method for four major journals from the institution database. Identified concepts were developed into a conceptual framework and validated using face to face semi-structured interviews involving ten key stakeholders. The findings are that stakeholder management and sustainability concepts are yet to embrace. This paper suggests a seven-step approach for project managers. (1 Embracing stakeholder management and sustainability concepts, (2 identify stakeholder approach and sustainable principle to adopt, (3 identify and classify project stakeholders, (4 engage stakeholders, (5 analyze stakeholders, (6 manage stakeholders and sustainable principle, (7 monitor and support process.

  5. [Pregnancy during residency].

    Maas, S M; van 't Hoff, B W; Rings, E H; van der Waals, F W; Büller, H A

    1992-12-19

    The number of female residents in the Netherlands has steadily increased in recent years. Due to the increased time on waiting lists to enter residency programmes and to the increased duration of training, female residents will be older during their residencies. This will probably result in an increased number of pregnancies during residencies. A questionnaire regarding pregnancy during residency was sent to 191 residents in two university hospitals in the Netherlands. The response rate was 74.3%. Fifty percent of the male and only 19% of the female residents had children. No negative effects of a pregnancy on their training were experienced or anticipated by the residents. However, a negative effect on the functioning of the department was expected. No formal provisions, like replacements were available and many solutions to replace pregnant colleagues depended on the flexibility of the colleagues. The wish to have children was high and equally distributed among male and female residents, 92% and 96%, resp. Given the difficulty to seek a permanent position and to have children after residency, the choice of many female residents will be to have their children during residency. This increase in number of pregnancies requires anticipation of the residency programme directors. They should take the lead in proposing adequate regulations. PMID:1470257

  6. pLS010 plasmid vector

    Lacks, Sanford A.; Balganesh, Tanjore S.

    1988-01-01

    Disclosed is recombinant plasmid pLS101, consisting essentially of a 2.0 Kb malM gene fragment ligated to a 4.4 Kb T.sub.c r DNA fragment, which is particularly useful for transforming Gram-positive bacteria. This plasmid contains at least four restriction sites suitable for inserting exogeneous gene sequences. Also disclosed is a method for plasmid isolation by penicillin selection, as well as processes for enrichment of recombinant plasmids in Gram-positive bacterial systems.

  7. Chromate resistance plasmid in Pseudomonas fluorescens.

    Bopp, L H; Chakrabarty, A M; Ehrlich, H. L.

    1983-01-01

    Chromate resistance of Pseudomonas fluorescens LB300, isolated from chromium-contaminated sediment in the upper Hudson River, was found to be plasmid specified. Loss of the plasmid (pLHB1) by spontaneous segregation or mitomycin C curing resulted in a simultaneous loss of chromate resistance. Subsequent transformation of such strains with purified pLHB1 plasmid DNA resulted in a simultaneous re-acquisition of the chromate resistance phenotype and the plasmid. When pLHB1 was transferred by con...

  8. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets.

    Kumar, Sudhir; Stecher, Glen; Tamura, Koichiro

    2016-07-01

    We present the latest version of the Molecular Evolutionary Genetics Analysis (Mega) software, which contains many sophisticated methods and tools for phylogenomics and phylomedicine. In this major upgrade, Mega has been optimized for use on 64-bit computing systems for analyzing larger datasets. Researchers can now explore and analyze tens of thousands of sequences in Mega The new version also provides an advanced wizard for building timetrees and includes a new functionality to automatically predict gene duplication events in gene family trees. The 64-bit Mega is made available in two interfaces: graphical and command line. The graphical user interface (GUI) is a native Microsoft Windows application that can also be used on Mac OS X. The command line Mega is available as native applications for Windows, Linux, and Mac OS X. They are intended for use in high-throughput and scripted analysis. Both versions are available from www.megasoftware.net free of charge. PMID:27004904

  9. ANALYSIS OF A MODEL OF PLASMID-BEARING, PLASMID-FREE COMPETITION IN A PULSED CHEMOSTAT

    XIANGYUN SHI; XINYU SONG; XUEYONG ZHOU

    2006-01-01

    We introduce and study a chemostat model with plasmid-bearing, plasmid-free competition and impulsive effect. According to the stability analysis of the boundary periodic solution, we obtain the invasion threshold of the plasmid-free organism and plasmid-bearing organism. Furthermore, by using standard techniques of bifurcation theory, we prove the system has a positive τ-periodic solution, which shows that the impulsive effect destroys the equilibria of the unforced continuous system and ini...

  10. Plasmid ColVBtrp maintenance in Erwinia carotovora.

    Schukin, N N

    1981-01-01

    Plasmid ColVBtrp maintenance in Erwinia carotovora cells was followed by measuring kinetics of elimination of plasmid genetic markers and loss of plasmid deoxyribonucleic acid. An E. carotovora mutant stably carrying plasmid ColVBtrp was isolated. Besides stable plasmid maintenance, the mutant showed altered sensitivity to male-specific phage MS2, sensitivity to drugs, and colony morphology.

  11. The expression of a plasmid-specified exported protein causes structural plasmid instability in Bacillus subtilis

    Cordes, C.; Meima, R; Twiest, B; Kazemier, B; Venema, G; vanDijl, JM; Bron, S

    1996-01-01

    The rolling-circle plasmid pGP1 was used to study the effects of the expression of a plasmid-specified exported protein on structural plasmid stability in Bacillus subtilis. pGP1 contains a fusion between the Bacillus licheniformis penP gene, encoding a C-terminally truncated penicillinase, and the

  12. Resident Characteristics Report

    Department of Housing and Urban Development — The Resident Characteristics Report summarizes general information about households who reside in Public Housing, or who receive Section 8 assistance. The report...

  13. Isolation of Plasmid DNA from Butyrivibrio fibrisolvens†

    Teather, Ronald M.

    1982-01-01

    A procedure based on successive precipitation of cell lysates with sodium dodecyl sulfate-NaCl and polyethylene glycol 6000 was developed which allows the isolation of plasmid DNA from Butyrivibrio fibrisolvens. A survey of B. fibrisolvens strains isolated from the bovine rumen showed that plasmids are a common feature of this species.

  14. Plasmid deoxyribonucleic acid replication in Streptomyces griseus.

    Xue, Y.; Zhuang, Z.; Zhu, Y.; Xu, Y.; Dong, K.

    1981-01-01

    A series of electron micrographs showing the presence of different molecular forms representing various replication stages of plasmid deoxyribonucleic acid from Streptomyces griseus was obtained. Based upon an analysis of these electron micrographs, a tentative model for plasmid deoxyribonucleic acid replication in S. griseus is proposed.

  15. Plasmid-mediated horizontal gene transfer is a coevolutionary process

    Harrison, Ellie; Brockhurst, Michael A

    2012-01-01

    Conjugative plasmids are key agents of horizontal gene transfer (HGT) that accelerate bacterial adaptation by vectoring ecologically important traits between strains and species. However, although many conjugative plasmids carry beneficial traits, all plasmids exert physiological costs-of-carriage on bacteria. The existence of conjugative plasmids, therefore, presents a paradox because non-beneficial plasmids should be lost to purifying selection, whereas beneficial genes carried on plasmids ...

  16. Conservation of plasmid DNA sequences in coronatine-producing pathovars of Pseudomonas syringae

    Bender, C.L.; Young, S.A. (Oklahoma State Univ., Stillwater (United States)); Mitchell, R.E. (Dept. of Scientific and Industrial Research, Auckland (New Zealand))

    1991-04-01

    In Pseudomonas syringae pv. tomato PT23.2, plasmid pPT23A (101 kb) is involved in synthesis of the phytotoxin coronatine. The physical characterization of mutations that abolished coronatine production indicated that at least 30 kb of pPT23A DNA are required for toxin synthesis. In the present study, {sup 32}P-labeled DNA fragments from the 30-kb region of pPT23A hybridized to plasmid DNAs from several coronatine-producing pathovars of P. syringae under conditions of high stringency. These experiments indicated that this region of pPT23A was strongly conserved in large plasmids (90 to 105 kb) that reside in P. syringae pv. atropurpurea, glycinea, and morsprunorum. The functional significance of the observed homology was demonstrated in marker-exchange experiments in which Tn5-inactivated sequences from the 30-kb region of pPT23A were used to mutate coronatine synthesis genes in the three heterologous pathovars. Physical characterization of the Tn5 insertions generated by marker exchange indicated that genes controlling coronatine synthesis in P. syringae pv. atropurpurea 1304, glycinea 4180, and morsprunorum 567 and 3714 were located on the large indigenous plasmids where homology was originally detected. Therefore, coronatine biosynthesis genes are strongly conserved in the plasmid DNAs of four producing pathovars, despite their disparate origins (California, Japan, New Zealand, Great Britain, and Italy).

  17. Conservation of plasmid DNA sequences in coronatine-producing pathovars of Pseudomonas syringae

    In Pseudomonas syringae pv. tomato PT23.2, plasmid pPT23A (101 kb) is involved in synthesis of the phytotoxin coronatine. The physical characterization of mutations that abolished coronatine production indicated that at least 30 kb of pPT23A DNA are required for toxin synthesis. In the present study, 32P-labeled DNA fragments from the 30-kb region of pPT23A hybridized to plasmid DNAs from several coronatine-producing pathovars of P. syringae under conditions of high stringency. These experiments indicated that this region of pPT23A was strongly conserved in large plasmids (90 to 105 kb) that reside in P. syringae pv. atropurpurea, glycinea, and morsprunorum. The functional significance of the observed homology was demonstrated in marker-exchange experiments in which Tn5-inactivated sequences from the 30-kb region of pPT23A were used to mutate coronatine synthesis genes in the three heterologous pathovars. Physical characterization of the Tn5 insertions generated by marker exchange indicated that genes controlling coronatine synthesis in P. syringae pv. atropurpurea 1304, glycinea 4180, and morsprunorum 567 and 3714 were located on the large indigenous plasmids where homology was originally detected. Therefore, coronatine biosynthesis genes are strongly conserved in the plasmid DNAs of four producing pathovars, despite their disparate origins (California, Japan, New Zealand, Great Britain, and Italy)

  18. Bifurcation Analysis of a Chemostat Model of Plasmid-Bearing and Plasmid-Free Competition with Pulsed Input

    Zhong Zhao; Baozhen Wang; Liuyong Pang; Ying Chen

    2014-01-01

    A chemostat model of plasmid-bearing and plasmid-free competition with pulsed input is proposed. The invasion threshold of the plasmid-bearing and plasmid-free organisms is obtained according to the stability of the boundary periodic solution. By use of standard techniques of bifurcation theory, the periodic oscillations in substrate, plasmid-bearing, and plasmid-free organisms are shown when some conditions are satisfied. Our results can be applied to control bioreactor aimed at producing co...

  19. Investigating fusion plasma instabilities in the Mega Amp Spherical Tokamak using mega electron volt proton emissions (invited)

    The proton detector (PD) measures 3 MeV proton yield distributions from deuterium-deuterium fusion reactions within the Mega Amp Spherical Tokamak (MAST). The PD’s compact four-channel system of collimated and individually oriented silicon detectors probes different regions of the plasma, detecting protons (with gyro radii large enough to be unconfined) leaving the plasma on curved trajectories during neutral beam injection. From first PD data obtained during plasma operation in 2013, proton production rates (up to several hundred kHz and 1 ms time resolution) during sawtooth events were compared to the corresponding MAST neutron camera data. Fitted proton emission profiles in the poloidal plane demonstrate the capabilities of this new system

  20. PcrA function in plasmid replication

    Chisty, L. T.

    2014-01-01

    PcrA is a DNA helicase involved in unwinding plasmi ds as a part of a complex in asymmetric rolling - circle replication of certain plasmids carrying antibiotic resistance genes. PcrA translocates on single stranded DNA by coupling ATP hydrolysis to movement on DNA. Initiator protein, RepD is required to nick supercoiled plasmid site - specifically and open an ssDNA stretch that PcrA can bind. The presence of RepD is needed throughout plasmid unwinding to maintain processivity. Using fluo...

  1. Complete sequences of four plasmids of Lactococcus lactis subsp. cremoris SK11 reveal extensive adaptation to the dairy environment.

    Siezen, Roland J; Renckens, Bernadet; van Swam, Iris; Peters, Sander; van Kranenburg, Richard; Kleerebezem, Michiel; de Vos, Willem M

    2005-12-01

    Lactococcus lactis strains are known to carry plasmids encoding industrially important traits. L. lactis subsp. cremoris SK11 is widely used by the dairy industry in cheese making. Its complete plasmid complement was sequenced and found to contain the plasmids pSK11A (10,372 bp), pSK11B (13,332 bp), pSK11L (47,165 bp), and pSK11P (75,814 bp). Six highly homologous repB-containing replicons were found, all belonging to the family of lactococcal theta-type replicons. Twenty-three complete insertion sequence elements segment the plasmids into numerous modules, many of which can be identified as functional units or containing functionally related genes. Plasmid-encoded functions previously known to reside on L. lactis SK11 plasmids were now mapped in detail, e.g., lactose utilization (lacR-lacABCDFEGX), the proteolytic system (prtM-prtP, pepO, pepF), and the oligopeptide permease system (oppDFBCA). Newly identified plasmid-encoded functions could facilitate the uptake of various cations, while the pabA and pabB genes could be essential for folate biosynthesis. A competitive advantage could be obtained by using the putative flavin adenine dinucleotide-dependent d-lactate dehydrogenase and oxalate:formate antiporter for enhanced ATP synthesis, while the activity of the predicted alpha-acetolactate decarboxylase may contribute to the formation of an additional electron sink. Various stress response proteins are plasmid encoded, which could enhance strain robustness. A substantial number of these "adaptation" genes have not been described before on L. lactis plasmids. Moreover, several genes were identified for the first time in L. lactis, possibly reflecting horizontal gene transfer. PMID:16332824

  2. A Functional oriT in the Ptw Plasmid of Burkholderia cenocepacia Can Be Recognized by the R388 Relaxase TrwC

    Fernández-González, Esther; Bakioui, Sawsane; Gomes, Margarida C.; O'Callaghan, David; Vergunst, Annette C.; Sangari, Félix J.; Llosa, Matxalen

    2016-01-01

    Burkholderia cenocepacia is both a plant pathogen and the cause of serious opportunistic infections, particularly in cystic fibrosis patients. B. cenocepacia K56-2 harbors a native plasmid named Ptw for its involvement in the Plant Tissue Watersoaking phenotype. Ptw has also been reported to be important for survival in human cells. Interestingly, the presence of PtwC, a homolog of the conjugative relaxase TrwC of plasmid R388, suggests a possible function for Ptw in conjugative DNA transfer. The ptw region includes Type IV Secretion System genes related to those of the F plasmid. However, genes in the adjacent region shared stronger homology with the R388 genes involved in conjugative DNA metabolism. This region included the putative relaxase ptwC, a putative coupling protein and accessory nicking protein, and a DNA segment with high number of inverted repeats and elevated AT content, suggesting a possible oriT. Although we were unable to detect conjugative transfer of the Ptw resident plasmid, we detected conjugal mobilization of a co-resident plasmid containing the ptw region homologous to R388, demonstrating the cloned ptw region contains an oriT. A similar plasmid lacking ptwC could not be mobilized, suggesting that the putative relaxase PtwC must act in cis on its oriT. Remarkably, we also detected mobilization of a plasmid containing the Ptw oriT by the R388 relaxase TrwC, yet we could not detect PtwC-mediated mobilization of an R388 oriT-containing plasmid. Our data unambiguously show that the Ptw plasmid harbors DNA transfer functions, and suggests the Ptw plasmid may play a dual role in horizontal DNA transfer and eukaryotic infection. PMID:27200362

  3. Enhanced Mission-Enabling Ultra-High Power Solar Array (Mega-ROSA EX) Project

    National Aeronautics and Space Administration — Mega-ROSA-EX is an enhanced, higher stiffness, higher sun-pointing accuracy, higher strength, higher specific power and even larger overall power / deployed size /...

  4. Mega Events and Urban Conflicts in Valencia, Spain: Contesting the New Urban Modernity

    Catherine Trudelle; Luis del Romero Renau

    2011-01-01

    The local and regional administrations of Valencia, Spain, have been working to position the city, which is the country's third largest in terms of population, as an international destination for luxury tourism through sports mega-events. Valencia's new image as a “mega-events city” has been criticized by citizens and urban-based movements on social and environmental grounds, resulting in various urban conflicts. The aim of this paper is to examine the causes and dynamics of these conflicts i...

  5. The Stabilization of High-rise Buildings : An Evaluation of the Tubed Mega Frame Concept

    Sandelin, Christian; Budajev, Evgenij

    2013-01-01

    Building tall has always been an expression of dreams, power and technical advancement. With the greatly increasing urbanization in recent years building tall has become a more viable option for office and residential housing. The Tubed mega frame concept tries to evolve the stabilizing systems of high-rise buildings with its mega frame around the buildings perimeter, created together with a new elevator system; the Articulated Funiculator. This thesis examines the effectiveness of the Tubed ...

  6. Wireless Laser Range Finder System for Vertical Displacement Monitoring of Mega-Trusses during Construction

    Hyo Seon Park; Sewook Son; Se Woon Choi; Yousok Kim

    2013-01-01

    As buildings become increasingly complex, construction monitoring using various sensors is urgently needed for both more systematic and accurate safety management and high-quality productivity in construction. In this study, a monitoring system that is composed of a laser displacement sensor (LDS) and a wireless sensor node was proposed and applied to an irregular building under construction. The subject building consists of large cross-sectional members, such as mega-columns, mega-trusses, a...

  7. The Kalimantan Border Oil Palm Mega-project

    Wakker, E. [AIDEnvironment, Amsterdam (Netherlands)

    2006-04-15

    A few years ago, the Indonesian government and sections of the palm oil industry united in the Indonesian Palm Oil Commission (IPOC) to undertake efforts to restore the atrocious public image that the palm oil industry had earned abroad for its role in the demise of Indonesia's tropical rainforests, the massive forest fires and haze in 1997-1998, and for the widespread conflicts between plantation companies and local communities. If IPOC succeeded in restoring the palm oil industry's image abroad, it was shattered again after June 2005 when the Indonesian Minister of Agriculture revealed details of a government plan to develop the world's largest oil palm plantation in a 5-10 kilometer band along the border of Kalimantan and Malaysia. To finance the USD 567 million plantation project, the Indonesian President and Chamber of Commerce and Industry (KADIN) had already met up with the Chinese government and private sector several times, resulting in Memoranda of Understanding between (among other) the Artha Graha and Sinar Mas groups from Indonesia and the Chinese CITIC group and Chinese Development Bank (CDB). The oil palm mega-project, launched in Indonesia under the banner of 'bringing prosperity, security and environmental protection to the Kalimantan border area', turned sour when a business plan developed by the Indonesian State Plantation Corporation (PTPN) began to circulate. This document contained a map that showed beyond doubt how the 1.8 million hectare oil palm project would trash the primary forests of three National Parks, cut through rugged slopes and mountains utterly unsuitable for oil palm cultivation and annihilate the customary rights land of the indigenous Dayak communities in the border area. This report describes what has come of the Kalimantan border oil palm mega-plan since it was announced, who is involved and what research, lobby and campaigning has led to so far. In particular, this study aims to inform civil

  8. The Kalimantan Border Oil Palm Mega-project

    A few years ago, the Indonesian government and sections of the palm oil industry united in the Indonesian Palm Oil Commission (IPOC) to undertake efforts to restore the atrocious public image that the palm oil industry had earned abroad for its role in the demise of Indonesia's tropical rainforests, the massive forest fires and haze in 1997-1998, and for the widespread conflicts between plantation companies and local communities. If IPOC succeeded in restoring the palm oil industry's image abroad, it was shattered again after June 2005 when the Indonesian Minister of Agriculture revealed details of a government plan to develop the world's largest oil palm plantation in a 5-10 kilometer band along the border of Kalimantan and Malaysia. To finance the USD 567 million plantation project, the Indonesian President and Chamber of Commerce and Industry (KADIN) had already met up with the Chinese government and private sector several times, resulting in Memoranda of Understanding between (among other) the Artha Graha and Sinar Mas groups from Indonesia and the Chinese CITIC group and Chinese Development Bank (CDB). The oil palm mega-project, launched in Indonesia under the banner of 'bringing prosperity, security and environmental protection to the Kalimantan border area', turned sour when a business plan developed by the Indonesian State Plantation Corporation (PTPN) began to circulate. This document contained a map that showed beyond doubt how the 1.8 million hectare oil palm project would trash the primary forests of three National Parks, cut through rugged slopes and mountains utterly unsuitable for oil palm cultivation and annihilate the customary rights land of the indigenous Dayak communities in the border area. This report describes what has come of the Kalimantan border oil palm mega-plan since it was announced, who is involved and what research, lobby and campaigning has led to so far. In particular, this study aims to inform civil society organizations, palm oil

  9. Plasmids as Tools for Containment.

    García, José L; Díaz, Eduardo

    2014-10-01

    Active containment systems are a major tool for reducing the uncertainty associated with the introduction of monocultures, genetically engineered or not, into target habitats for a large number of biotechnological applications (e.g., bioremediation, bioleaching, biopesticides, biofuels, biotransformations, live vaccines, etc.). While biological containment reduces the survival of the introduced organism outside the target habitat and/or upon completion of the projected task, gene containment strategies reduce the lateral spread of the key genetic determinants to indigenous microorganisms. In fundamental research, suicide circuits become relevant tools to address the role of gene transfer, mainly plasmid transfer, in evolution and how this transfer contributes to genome plasticity and to the rapid adaptation of microbial communities to environmental changes. Many lethal functions and regulatory circuits have been used and combined to design efficient containment systems. As many new genomes are being sequenced, novel lethal genes and regulatory elements are available, e.g., new toxin-antitoxin modules, and they could be used to increase further the current containment efficiencies and to expand containment to other organisms. Although the current containment systems can increase the predictability of genetically modified organisms in the environment, containment will never be absolute, due to the existence of mutations that lead to the appearance of surviving subpopulations. In this sense, orthogonal systems (xenobiology) appear to be the solution for setting a functional genetic firewall that will allow absolute containment of recombinant organisms. PMID:26104372

  10. Enhancement of plasmid-mediated gene therapy for muscular dystrophy by directed plasmid integration

    Bertoni, Carmen; Jarrahian, Sohail; Wheeler, Thurman M.; LI, YINING; Olivares, Eric C.; Michele P Calos; Rando, Thomas A.

    2005-01-01

    Plasmid-mediated gene therapy can restore dystrophin expression in skeletal muscle in the mdx mouse, a model of Duchenne muscular dystrophy. However, sufficient long-term expression and distribution of dystrophin remain a hurdle for translating this technology into a viable treatment for Duchenne muscular dystrophy. To improve plasmid-mediated gene therapy for muscle diseases, we studied the effects of targeted plasmid integration using a phage integrase (φC31) that can mediate the integratio...

  11. Plasmid DNA fermentation strategies: influence on plasmid stability and cell physiology.

    Silva, Filomena; Queiroz, João A; Domingues, Fernanda C

    2012-03-01

    In order to provide sufficient pharmaceutical-grade plasmid DNA material, it is essential to gain a comprehensive knowledge of the bioprocesses involved; so, the development of protocols and techniques that allow a fast monitoring of process performance is a valuable tool for bioprocess design. Regarding plasmid DNA production, the metabolic stress of the host strain as well as plasmid stability have been identified as two of the key parameters that greatly influence plasmid DNA yields. The present work describes the impact of batch and fed-batch fermentations using different C/N ratios and different feeding profiles on cell physiology and plasmid stability, investigating the potential of these two monitoring techniques as valuable tools for bioprocess development and design. The results obtained in batch fermentations showed that plasmid copy number values suffered a pronounced increase at the end of almost all fermentation conditions tested. Regarding fed-batch fermentations, the strategies with exponential feeding profiles, in contrast with those with constant feeding, showed higher biomass and plasmid yields, the maximum values obtained for these two parameters being 95.64 OD(600) and 344.3 mg plasmid DNA (pDNA)/L, respectively, when using an exponential feed rate of 0.2 h(-1). Despite the results obtained, cell physiology and plasmid stability monitoring revealed that, although higher pDNA overall yields were obtained, this fermentation exhibited lower plasmid stability and percentage of viable cells. In conclusion, this study allowed clarifying the bioprocess performance based on cell physiology and plasmid stability assessment, allowing improvement of the overall process and not only plasmid DNA yield and cell growth. PMID:22089386

  12. Conjugal mobilization of the mega element carrying mef(E) from Streptococcus salivarius to Streptococcus pneumoniae.

    Santagati, Maria; Lupo, Agnese; Scillato, Marina; Di Martino, Andrea; Stefani, Stefania

    2009-01-01

    We report the isolation and characterization of an unusual strain of Streptococcus salivarius, 3C30, displaying both the macrolide-lincosamide-streptogramin B and the tetracycline resistance phenotypes. It harbours the mef(E), erm(B), and tet(M) genes carried by different genetic elements. The genetic element carrying mef(E), named mega, was investigated by long PCR and sequencing, while the presence of the Tn3872-like element, carrying tet(M) and erm(B), was demonstrated by sequencing of both the int-xis-Tn and the fragment between the two resistance genes. In strain 3C30 the mega element is 5388 bp in size and its nucleotide sequence is identical to that of the element described previously in S. salivarius, with the exception of a 912 bp deletion at the left end. The composite Tn3872-like element appeared to be nonconjugative while the mega element was transferred by conjugation to Streptococcus pneumoniae. It was, however, impossible to transfer it again from these transconjugants to other strains. In addition, only in the 3C30 strain did mega form circular structures, as identified by real-time PCR. In conclusion, we found a clinical strain of S. salivarius carrying both mega and Tn3872-like genetic elements. Mega is transferable by conjugation to S. pneumoniae but it is not transferable again from the transconjugants, suggesting a possible mobilization by recombinases of the coresident Tn3872-like transposon. PMID:19025575

  13. Elimination of multicopy plasmid R6K by bleomycin.

    Attfield, P V; Pinney, R. J.

    1985-01-01

    Bleomycin eliminated multicopy plasmid R6K from growing cells of Escherichia coli AB1157 but failed to cure either of the low-copy plasmids R1 or R46. Measurements of R6K-encoded beta-lactamase and of covalently closed plasmid DNA indicated that the drug causes a progressive reduction in plasmid copy number.

  14. Behavior of IncQ Plasmids in Agrobacterium tumefaciens

    Hille, Jacques; Schilperoort, Rob

    1981-01-01

    Inc-Q plasmids were introduced into Agrobacterium tumefuciens, by mobilization from Escherichia coli with an Inc-P plasmid, or by transformation with purified plasmid DNA. It was found that they were stably maintained. The presence of an Inc-Q plasmid did not influence tumorigenicity. These results

  15. Curing of plasmid pXO1 from Bacillus anthracis using plasmid incompatibility.

    Xiankai Liu

    Full Text Available The large plasmid pXO1 encoding the anthrax toxin is important for the virulence of Bacillus anthracis. It is essential to cure pXO1 from B. anthracis to evaluate its role in the pathogenesis of anthrax infection. Because conventional methods for curing plasmids (e.g., curing agents or growth at elevated temperatures can induce mutations in the host chromosomal DNA, we developed a specific and reliable method to eliminate pXO1 from B. anthracis using plasmid incompatibility. Three putative replication origins of pXO1 were inserted into a temperature-sensitive plasmid to generate three incompatible plasmids. One of the three plasmids successfully eliminated the large plasmid pXO1 from B. anthracis vaccine strain A16R and wild type strain A16. These findings provided additional information about the replication/partitioning of pXO1 and demonstrated that introducing a small incompatible plasmid can generate plasmid-cured strains of B. anthracis without inducing spontaneous mutations in the host chromosome.

  16. Positive epistasis between co-infecting plasmids promotes plasmid survival in bacterial populations

    San Millan, Alvaro; Heilbron, Karl; MacLean, R. Craig

    2014-01-01

    Plasmids have a key role in the horizontal transfer of genes among bacteria. Although plasmids are catalysts for bacterial evolution, it is challenging to understand how they can persist in bacterial populations over the long term because of the burden they impose on their hosts (the ‘plasmid paradox'). This paradox is especially perplexing in the case of ‘small' plasmids, which are unable to self-transfer by conjugation. Here, for the first time, we investigate how interactions between co-in...

  17. Mini-F plasmid genes that couple host cell division to plasmid proliferation.

    Ogura, T; Hiraga, S

    1983-01-01

    A mechanism for stable maintenance of plasmids, besides the replication and partition mechanisms, has been found to be specified by genes of a mini-F plasmid. An oriC plasmid carrying both a mini-F segment necessary for partition [coordinates 46.4-49.4 kilobase pairs (kb) on the F map] and another segment (42.9-43.6 kb), designated ccd (coupled cell division), is more stably maintained than are oriC plasmids carrying only the partition segment; the stability is comparable to that of the paren...

  18. Adaptive Plasmid Evolution Results in Host-Range Expansion of a Broad-Host-Range Plasmid

    De Gelder, Leen; Williams, Julia J.; Ponciano, José M; Sota, Masahiro; Eva M. Top

    2008-01-01

    Little is known about the range of hosts in which broad-host-range (BHR) plasmids can persist in the absence of selection for plasmid-encoded traits, and whether this “long-term host range” can evolve over time. Previously, the BHR multidrug resistance plasmid pB10 was shown to be highly unstable in Stenotrophomonas maltophilia P21 and Pseudomonas putida H2. To investigate whether this plasmid can adapt to such unfavorable hosts, we performed evolution experiments wherein pB10 was maintained ...

  19. Plasmid and chromosome segregation in prokaryotes

    Møller-Jensen, Jakob; Bugge Jensen, Rasmus; Gerdes, Kenn

    2000-01-01

    Recent major advances in the understanding of prokaryotic DNA segregation have been achieved by using fluorescence microscopy to visualize the localization of cellular components. Plasmids and bacterial chromosomes are partitioned in a highly dynamic fashion, suggesting the presence of a mitotic......-like apparatus in prokaryotes. The identification of chromosomal homologues of the well-characterized plasmid partitioning genes indicates that there could be a general mechanism of bacterial DNA partitioning. Udgivelsesdato: July 1...

  20. Characterization of a Haemophilus ducreyi mobilizing plasmid.

    McNicol, P J; Albritton, W L; Ronald, A R

    1986-01-01

    The OriV site of Haemophilus ducreyi mobilizing plasmid pHD147, determined by replication in Escherichia coli polA, is located close to the OriT site. The OriT site, located by recombination-proficient and -deficient cells, and the OriV site map in a region of pHD147 homologous to the beta-lactamase-specifying plasmids of H. ducreyi and Neisseria gonorrhoeae.

  1. Plasmid Evolution and Interaction between the Plasmid Addiction Stability Systems of Two Related Broad-Host-Range IncQ-Like Plasmids

    Deane, Shelly M.; Rawlings, Douglas E

    2004-01-01

    Plasmid pTC-F14 contains a plasmid stability system called pas (plasmid addiction system), which consists of two proteins, a PasA antitoxin and a PasB toxin. This system is closely related to the pas of plasmid pTF-FC2 (81 and 72% amino acid identity for PasA and PasB, respectively) except that the pas of pTF-FC2 contains a third protein, PasC. As both pTC-F14 and pTF-FC2 are highly promiscuous broad-host-range plasmids isolated from bacteria that share a similar ecological niche, the plasmid...

  2. Beyond the MEGA BORG: The future of cooperative damage assessments

    Prespill planning and coordination are the major requirements of industry and government to ensure future cooperative natural resource damage assessments (NRDA). Since oil spills most often occur without warning, both parties must be prepared prior to an oil spill into the environment. Federal and State co-trustees should enter into a Memorandum of Understanding (MOU) to ensure cooperation and coordination among trustees in all NRDA activities. The MOU should provide the framework whereby each trustee's resource interests can be represented. Ideally, MOUs should be prepared during the prespill planning process, although the MEGA BORG MOU was prepared soon after the actual discharge. The establishment of co-trustee working groups will foster the positive working relationships necessary in cooperative NRDAs. The trustees should identify all potential co-trustees in a given area and agree on a mechanism for trustee notification. Criteria for the rapid designation of the Lead Administrative Trustee and approaches for coordination with response agencies and potential responsible parties (PRPs) need to be developed. Most importantly, the trustees should identify PRPs and invite their participation in the prespill planning process

  3. Mega-mergers in the pharmaceutical industry. In whose interests?

    Po, A L

    1998-10-01

    Companies merge to achieve economies of scale. In an industry such as the pharmaceutical industry which relies on a high level of investment in research and development, such mergers appear rational. However, it is not at all obvious that a higher level of investment by a smaller number of firms will necessarily lead to an increased rate of genuine innovations. There is a risk that conflicts of interest and the pursuit of short term gains may encourage more mergers than is optimal for the industry. The impact of mega-mergers in the pharmaceutical industry on research output, employees, shareholders, financial advisers, managers and patients is discussed. A healthy pharmaceutical industry, able to invest the necessary resources in the development of innovative medicines is in the interest of patients and shareholders alike. Over-concentration may interfere with innovative activity and lead to monopolistic power. Close scrutiny of merger activity is important but in a deregulated world, governments may have little power to act. In any case, a drug-specific monopolistic industry may be beneficial to some countries which may therefore be reluctant to act in the interest of the world as a single community in search of more effective medicines. PMID:10344903

  4. Mega-History and the 21st century singularity puzzle

    Akop P. Nazaretyan

    2015-06-01

    Full Text Available A series of calculations carried out independently by the Australian, Russian and American re- searchers have demonstrated that a crucial global polyfurcation is expected near the middle of the 21st century. This result is drawn by extrapolating into the future the logarithmic acceleration law, which involves the phase transitions in the evolution of biosphere and anthroposphere. The paper investigates the palliatives of the planetary civilization beyond the big evolutionary Singularity in the context of Mega-history and complexity theory worldviews. It gives the mathematical deduction a universal ground and besides, helps involve some recent discoveries in psychology and cultural anthropology to tracing the forecasting attractors and scenarios. The destiny of the Earth (as well as any other planetary civilization may conclusively depend on whether or not the intellectual ac- tor succeeds in developing his inner regulation to balance the potentially unlimited developments in technological power. Particularly, this includes overcoming the macro-group identities, religious and quasi-religious ideologies, which always suggest a friend-or-foe discrimination matrix.

  5. FCC resid processing

    This paper narrows the definition of reside processor to those FCC's which run feedstocks containing 1 wt.% or more Conradson carbon residue. With this new definition, the resid survey is revisited to see if any new conclusions can be drawn from the data. The authors break down the numbers and geography of resid processors, feed types, operating variables, and yields. After examining the state of resid processing in the FCC, the paper focuses on the design of cracking catalysts for handling resid feeds. There are important considerations involved in processing resid including high levels of contaminants such as vanadium and nickel, the impact on heat balance, and diffusion effects. Catalysts can be designed to improve the profitability of a resid operation. A clear picture of the roles of zeolite and matrix is presented, along with a discussion of the different types of zeoliters which are commonly used. The paper demonstrates how zeolite and matrix are best combined to meet objectives within a given set of constraints while processing resid

  6. Plasmids spread very fast in heterogeneous bacterial communities.

    Dionisio, Francisco; Matic, Ivan; Radman, Miroslav; Rodrigues, Olivia R; Taddei, François

    2002-01-01

    Conjugative plasmids can mediate gene transfer between bacterial taxa in diverse environments. The ability to donate the F-type conjugative plasmid R1 greatly varies among enteric bacteria due to the interaction of the system that represses sex-pili formations (products of finOP) of plasmids already harbored by a bacterial strain with those of the R1 plasmid. The presence of efficient donors in heterogeneous bacterial populations can accelerate plasmid transfer and can spread by several order...

  7. Plasmid Segregation: Spatial Awareness at the Molecular Level

    Møller-Jensen, Jakob; Gerdes, Kenn

    2007-01-01

    In bacteria, low-copy number plasmids ensure their stable inheritance by partition loci (par), which actively distribute plasmid replicates to each side of the cell division plane. Using time-lapse fluorescence microscopic tracking of segregating plasmid molecules, a new study provides novel...... insight into the workings of the par system from Escherichia coli plasmid R1. Despite its relative simplicity, the plasmid partition spindle shares characteristics with the mitotic machinery of eukaryotic cells....

  8. Mobilization of Thiobacillus ferrooxidans plasmids among Escherichia coli strains.

    Rawlings, D. E.; Woods, D R

    1985-01-01

    Nonconjugative Thiobacillus ferrooxidans plasmids were mobilized at high frequencies among Escherichia coli strains by the IncP plasmid RP4 and at low frequencies by the IncN plasmid R46, but not by the IncW plasmid pSa. The mobilization region of a nonconjugative T. ferrooxidans plasmid was located on a 5.3-kilobase T. ferrooxidans DNA fragment.

  9. Relationship between plasmid content and auxotype in Neisseria gonorrhoeae isolates.

    Dillon, J R; Pauzé, M

    1981-01-01

    One hundred and forty strains of Neisseria gonorrhoeae, representing 12 different auxotype groups, were examined for differences in plasmid content. Most auxotype groups harbored a phenotypically cryptic 2,6-megadalton plasmid; a few groups also carried a 24.5-megadalton plasmid which has been previously characterized as a transfer plasmid. However, isolates of the proline-, citrulline-, and uracil-requiring (PCU-) auxotype were consistently free of plasmids. The correlation between auxotype ...

  10. Plasmid stability and maintenance of copy number using natural marker

    Hamzah Basil Mohammed; Sudhakar Malla

    2015-01-01

    Present study was conducted to study the plasmid stability with the help of natural plasmid isolated from the bacteria which lodges the ink gland of the sea squid and emits bioluminescence. Isolated bacterial strain was identified by using 16srRNA sequencing and its plasmid DNA was used for the experimental studies. The plasmid is found to be responsible for the bioluminescence. The stability of this plasmid was studied in shake flask method using the different sugar sources (Gluc...

  11. Isolation of a Pseudomonas Stutzeri strain that degrades1, 2, 4-trichlorobenzene and characterization of its degradative plasmid

    Lei SONG; Hui WANG; Hanchang SHI; Hongying HU

    2008-01-01

    The genetic information encoding metabolic pathways for xenobiotic compounds in bacteria often resides on catabolic plasmids. The aim of the present work was to know the location of the genes for degrading 1, 2, 4-trichlorobenzen. In this paper a 1, 2, 4-trichlorobenzene-degrading strain THSL-1 was isolated from the soil of Tianjin Chemical Plant using 1, 2, 4-trichlorobenzene as the sole carbon source. The strain was identified as Pseudomonas stutzeri through morphologic survey and 16S rDNA sequence determination. A plasmid was discovered from strain THSL-1 by using the alkali lysis method. When the plasmid was transformed into E. coli. JM109 by the CaCl2 method, the transformant could grow using 1, 2, 4-trichlorobenzene as the sole carbon source and had the degradation function of 1, 2, 4-trichlorobenzene. Therefore, it could be deemed that the plasmid carried the degradative genes of 1, 2, 4-trichlorobenzene. The average size of the plasmid was finally determined to be 40.2 Kb using selectively three kinds of restricted inscribed enzymes (HindIII, BamHI, and XholI) for single cutting and double cutting the plasmid pTHSL-1, respectively.

  12. Inducible Escherichia coli fermentation for increased plasmid DNA production.

    Carnes, Aaron E; Hodgson, Clague P; Williams, James A

    2006-11-01

    Bacterial plasmids are the vectors of choice for DNA vaccines and short-term gene therapeutics. Growing plasmid DNA by microbial (Escherichia coli) fermentation is usually combined with alkaline lysis/chromatography methods of purification. To date, typical plasmid fermentation media and processes result in yields of 100-250 mg of plasmid DNA/l of culture medium, using standard high-copy pUC origin-containing plasmids. In order to address this initial and yield-limiting upstream step, we identified novel fermentation control parameters for fed-batch fermentation. The resulting fermentation strategies significantly increased specific plasmid yield with respect to cell mass while enhancing plasmid integrity and maintaining supercoiled DNA content. Fed-batch fermentation yield exceeding 1000 mg of plasmid DNA/l was obtained after reduction of plasmid-mediated metabolic burden during growth, and yields up to 1500 mg of plasmid DNA/l have been achieved with optimized plasmid backbones. Interestingly, by inducing high plasmid levels after sufficient biomass accumulation at low temperature and restricted growth, cells were able to tolerate significantly higher plasmid quantities than cells grown by conventional processes. This 5-10-fold increase in plasmid yield dramatically decreases plasmid manufacturing costs and improves the effectiveness of downstream purification by reducing the fraction of impurities. PMID:16819941

  13. Conservation of Salmonella typhimurium virulence plasmid maintenance regions among Salmonella serovars as a basis for plasmid curing.

    Tinge, S A; Curtiss, R

    1990-01-01

    The association of large plasmids with virulence in invasive Salmonella serovars has led to a number of studies designed to uncover the role of these plasmids in virulence. This study addresses two aspects of virulence-associated plasmids. The first is the distribution of the replication and maintenance regions among the plasmids of different Salmonella serovars, and the second is the use of the conserved virulence plasmid par region to provide a rapid method for eliminating the virulence pla...

  14. BioShuttle-mediated Plasmid Transfer

    Klaus Braun, Leonie von Brasch, Ruediger Pipkorn, Volker Ehemann, Juergen Jenne, Herbert Spring, Juergen Debus, Bernd Didinger, Werner Rittgen, Waldemar Waldeck

    2007-01-01

    Full Text Available An efficient gene transfer into target tissues and cells is needed for safe and effective treatment of genetic diseases like cancer. In this paper, we describe the development of a transport system and show its ability for transporting plasmids. This non-viral peptide-based BioShuttle-mediated transfer system consists of a nuclear localization address sequence realizing the delivery of the plasmid phNIS-IRES-EGFP coding for two independent reporter genes into nuclei of HeLa cells. The quantification of the transfer efficiency was achieved by measurements of the sodium iodide symporter activity. EGFP gene expression was measured with Confocal Laser Scanning Microscopy and quantified with biostatistical methods by analysis of the frequency of the amplitude distribution in the CLSM images. The results demonstrate that the “BioShuttle”-Technology is an appropriate tool for an effective transfer of genetic material carried by a plasmid.

  15. BioShuttle-mediated Plasmid Transfer

    Braun, Klaus; von Brasch, Leonie; Pipkorn, Ruediger; Ehemann, Volker; Jenne, Juergen; Spring, Herbert; Debus, Juergen; Didinger, Bernd; Rittgen, Werner; Waldeck, Waldemar

    2007-01-01

    An efficient gene transfer into target tissues and cells is needed for safe and effective treatment of genetic diseases like cancer. In this paper, we describe the development of a transport system and show its ability for transporting plasmids. This non-viral peptide-based BioShuttle-mediated transfer system consists of a nuclear localization address sequence realizing the delivery of the plasmid phNIS-IRES-EGFP coding for two independent reporter genes into nuclei of HeLa cells. The quantification of the transfer efficiency was achieved by measurements of the sodium iodide symporter activity. EGFP gene expression was measured with Confocal Laser Scanning Microscopy and quantified with biostatistical methods by analysis of the frequency of the amplitude distribution in the CLSM images. The results demonstrate that the “BioShuttle”-Technology is an appropriate tool for an effective transfer of genetic material carried by a plasmid. PMID:18026568

  16. Burnout among Dutch medical residents

    Prins, J.T.; Hoekstra-Weebers, J.E.; van de Wiel, H.B.; Gazendam-Donofrio, S.M.; Sprangers, F.; Jaspers, F.C.; van der Heijden, F.M.

    2007-01-01

    We examined levels of burnout and relationships between burnout, gender, age, years in training, and medical specialty in 158 medical residents working at the University Medical Center Groningen, the Netherlands. Thirteen percent of the residents met the criteria for burnout, with the highest percentage of burnout cases among medical residents in Psychiatry. Significantly more male residents than female residents suffered from severe burnout. Medical residents reported significantly lower mea...

  17. Electrotransfer of Plasmid Vector DNA into Muscle

    Miyazaki, Satsuki; Miyazaki, Jun-Ichi

    Wolff et al. (1990) first reported that plasmid DNA injected into skeletal muscle is taken up by muscle cells and the genes in the plasmid are expressed for more than two months thereafter, although the transfected DNA does not usually undergo chromosomal integration (Wolff et al., 1991, 1992). However, the relatively low expression levels attained by this method have hampered its applications for uses other than as a DNA vaccine (Davis et al., 1995). There are a number of reports analyzing the conditions that affect the efficiency of gene transfer by intramuscular DNA injection and assessing the fine structures of expression plasmid vectors that may affect expression levels (Davis et al., 1993; Liang et al., 1996; Norman et al., 1997). Furthermore, various attempts were done to improve the efficiency of gene transfer by intramus cular DNA injection. Consequently, regenerating muscle was shown to produce 80-fold or more protein than did normal muscle, following injection of an expression plas-mid. Muscle regeneration was induced by treatment with cardiotoxin or bupivacaine (Wells, 1993; Vitadello et al., 1994). We previously demonstrated that by combining a strong promoter and bupivacaine pretreatment intramuscular injection of an IL-5 expression plasmid results in IL-5 production in muscle at a level sufficient to induce marked proliferation of eosinophils in the bone marrow and eosinophil infiltration of various organs (Tokui et al., 1997). It was also reported that a single intramuscular injection of an erythropoietin expression plasmid produced physiologically significant elevations in serum erythropoietin levels and increased hematocrits in adult mice (Tripathy et al., 1996). Hematocrits in these animals remained elevated at >60% for at least 90 days after a single injection. However, improvements to this method have not been sufficient to extend its applications including clinical use.

  18. BioShuttle-mediated Plasmid Transfer

    Braun, Klaus; von Brasch, Leonie; Pipkorn, Ruediger; Ehemann, Volker; Jenne, Juergen; Spring, Herbert; Debus, Juergen; Didinger, Bernd; Rittgen, Werner; Waldeck, Waldemar

    2007-01-01

    An efficient gene transfer into target tissues and cells is needed for safe and effective treatment of genetic diseases like cancer. In this paper, we describe the development of a transport system and show its ability for transporting plasmids. This non-viral peptide-based BioShuttle-mediated transfer system consists of a nuclear localization address sequence realizing the delivery of the plasmid phNIS-IRES-EGFP coding for two independent reporter genes into nuclei of HeLa cells. The quantif...

  19. Improvement of Performance of MegaBlast Algorithm for DNA Sequence Alignment

    Guang-Ming Tan; Lin Xu; Dong-Bo Bu; Sheng-Zhong Feng; Ning-Hui Sun

    2006-01-01

    MegaBlast is one of the most important programs in NCBI BLAST (Basic Local Alignment Search Tool)toolkits. However, MegaBlast is computation and I/O intensive. It consumes a great deal of memory which is proportional to the size of the query sequences set and subject (database) sequences set of product. This paper proposes a new strategy for optimizing MegaBlast. The new strategy exchanges the query and subject sequences sets, and builds a hash table based on new subject sequences. It overlaps I/O with computation, shortens the overall time and reduces the cost of memory,since the memory here is only proportional to the size of subject sequences set. The optimized algorithm is suitable to be parallelized in cluster systems. The parallel algorithm uses query segmentation method. As our experiments shown, the parallel program which is implemented with MPI has fine scalability.

  20. Competition between Plasmid-Bearing and Plasmid-Free Organisms in a Chemostat with Pulsed Input and Washout

    Sanling Yuan

    2009-01-01

    Full Text Available We consider a model of competition between plasmid-bearing and plasmid-free organisms in the chemostat with pulsed input and washout. We investigate the subsystem with nutrient and plasmid-free organism and study the stability of the boundary periodic solutions, which are the boundary periodic solutions of the system. The stability analysis of the boundary periodic solution yields the invasion threshold of the plasmid-bearing organism. By using the standard techniques of bifurcation theory, we prove that above this threshold there are periodic oscillations in substrate, plasmid-free, and plasmid-bearing organisms. Numerical simulations are carried out to illustrate our results.

  1. Shear viscosity of shocked metals at mega-bar pressures

    Liu, Fu-Sheng

    2013-06-01

    Viscosity of metals at high pressures and temperatures has been one of the most concerned problems in weapon physics and geophysics, e.g., the shear viscosity coefficients of substances in earth's mantle and earth's core at mega-bar pressures are needed for understanding the core mantle convection in deep earth. But the experimental data is very scarce because the conventional measurement methods can hardly be applied to such compression conditions [1]. In this talk, the principle of small-disturbance perturbation method [2] is re-investigated based on both the analytic solution and the numerical solution of the two-dimentional shock flow of sinusoidal distubance on front. In numerical solution, the real viscosity, which governs the flow behind the shock front and the perturbation damping feature, and the artificial viscosity, whick controls the numerical oscillation, separately treated. The relation between the viscosity of flow and the damping features of perturbation amplitude is quantitatively established for the loading situations of Sakharov's [3] and a flyer-impact situation with a finite disturbance. The later is the theoretical basis to develop a new experimental method, called the flyer-impact small-disturbance method [4]. In the flyer-impact small-disturbance method, the two-stage light-gas gun is used to launch a metal flyer. When the flyer directly impacts on the wedge-shaped sample with a sinusoidal surface, a two-dimensional shock flow of sinusoidal distubance on its front is generated. The amplitude of disturbance and its dependance with propagation distance is measured by use of an electric pin-array probe or a fibre-array probe. Correspondingly, the solution of the flow is given by numerically solving the hydrodynamic equations by the finite difference technique to find out the quantative correlations among the amplitude decay, the initial distribution of flow, the amplitude of initial disturbance, the shear viscosity of the flow, and the material

  2. Plasmid transduction by Bacillus subtilis bacteriophage SPP1: effects of DNA homology between plasmid and bacteriophage.

    Deichelbohrer, I; Alonso, J.C.; Lüder, G; Trautner, T A

    1985-01-01

    Any SPP1 DNA restriction fragment cloned into Bacillus subtilis plasmid pC194 or pUB110 increased the transduction frequency of the plasmid by SPP1 100- to 1,000-fold over the transduction level of the plasmid alone. This increment was observed irrespective of whether a fragment contained the SPP1 packaging origin (pac). Furthermore, an SPP1 derivative into whose genome pC194 DNA had been integrated transduced pC194 DNA with a greatly enhanced frequency. Transduction enhancement mediated by D...

  3. Plasmid maintenance functions of the large virulence plasmid of Shigella flexneri.

    Radnedge, L; Davis, M. A.; Youngren, B; Austin, S. J.

    1997-01-01

    The large virulence plasmid pMYSH6000 of Shigella flexneri contains a replicon and a plasmid maintenance stability determinant (Stb) on adjacent SalI fragments. The presence of a RepFIIA replicon on the SalI C fragment was confirmed, and the complete sequence of the adjacent SalI O fragment was determined. It shows homology to part of the transfer (tra) operon of the F plasmid. Stb stabilizes a partition-defective P1 miniplasmid in Escherichia coli. A 1.1-kb region containing a homolog of the...

  4. Plasmid transfer and plasmid-mediated genetic exchange in Brucella abortus.

    Rigby, C E; Fraser, A.D.

    1989-01-01

    Naturally-occurring plasmids and gene transfer mechanisms have not yet been reported in brucellae. Here we show that Brucella abortus is capable of maintaining and transferring the broad-host-range plasmids pTH10 (IncP), pSa (IncW) and R751 (IncP), and describe pTH10-mediated transfer of B. abortus chromosomal genes to Escherichia coli. All three plasmids transferred by conjugation from E. coli to B. abortus S19, and from B. abortus S19 to B. abortus 292 (biovar 4). They were stably maintaine...

  5. Challenges to publishing pharmacy resident research projects from the perspectives of residency program directors and residents

    Olson KL; Irwin AN; Joline BR; Witt DM; Patel RJ

    2013-01-01

    Objective: To identify barriers to completing and publishing pharmacy residency research projects from the perspective of program directors and former residents.Methods: This was a cross-sectional survey of pharmacy residency program directors and former post-graduate year one and two residents. Directors of pharmacy residency programs whose residents present their projects at the Western States Conference (n=216) were invited to complete an online survey and asked to forward the survey to fo...

  6. Identification, characterization and preliminary X-ray diffraction analysis of the rolling-circle replication initiator protein from plasmid pSTK1

    A proteolytically stable fragment of a plasmid replication initiation protein from the thermophile G. stearothermophilus has been biochemically characterized, crystallized and diffraction data collected to a resolution of 2.5 Å. Antibiotic resistance in bacterial pathogens poses an ever-increasing risk to human health. In antibiotic-resistant strains of Staphylococcus aureus this resistance often resides in extra-chromosomal plasmids, such as those of the pT181 family, which replicate via a rolling-circle mechanism mediated by a plasmid-encoded replication initiation protein. Currently, there is no structural information available for the pT181-family Rep proteins. Here, the crystallization of a catalytically active fragment of a homologous replication initiation protein from the thermophile Geobacillus stearothermophilus responsible for the replication of plasmid pSTK1 is reported. Crystals of the RepSTK1 fragment diffracted to a resolution of 2.5 Å and belonged to space group P212121

  7. Changes in medicine: residency

    Robbins RA

    2011-07-01

    Full Text Available No abstract available. Article truncated at 150 words. The most important time in a physician’s educational development is residency, especially the first year. However, residency work and responsibility have come under the scrutiny of a host of agencies and bureaucracies, and therefore, is rapidly changing. Most important in the alphabet soup of regulatory agencies is the Accreditation Council for Graduate Medical Education (ACGME which accredits residencies and ultimately makes the governing rules.Resident work hours have received much attention and are clearly decreasing. However, the decline in work hours began in the 1970’s before the present political push to decrease work hours. The residency I entered in 1976 had every third night call during the first year resident’s 6-9 months on general medicine or wards. It had changed from every other night the year before. On wards, we normally were in the hospital for our 24 hours of call and followed this with a 10-12 hour day before …

  8. Mega sporting events: A poisoned chalice or a new dawn for low- and middle-income

    Mark Tomlinson

    2011-06-01

    Full Text Available Mega sporting events in South Africa, which has the largest number of HIV-positive people in the world, and India, with 1.8 million deaths of children under 5 each year and 52 million stunted children, raise questions about the effective and, as importantly, the moral imperative of spending billions of dollars to host a sporting event. From a health perspective, selling alcohol and debt tarnishes further the notion of any intangible benefits of mega events to low- and middle-income countries.

  9. Development of large area resistive electrodes for ATLAS NSW MicroMEGAS

    Ochi, Atsuhiko; The ATLAS collaboration

    2015-01-01

    MicroMegas with resistive anode will be used for the NSW upgrade of the ATLAS experiment at LHC. The resistive electrode is one of key technology for MPGDs to prevent sparks. Large area resistive electrodes for the MM have been developed using two different technology; screen printing and carbon sputtering. Maximum size of each resistive foil is 45cm x 220cm with printed pattern of 425 micron pitch strips. Those technologies are also suitable to mass production. The prototypes of series production model have been produced successfully. We will report the development and production status and test results of the resistive MicroMegas.

  10. Simple method for identification of plasmid-coded proteins

    Proteins encoded by plasmid DNA are specifically labeled in uv-irradiated cells of Escherichia coli carrying recA and uvrA mutations because extensive degradation of the chromosome DNA occurs concurrently with amplification of plasmid DNA

  11. Plasmid maintenance and protein overproduction in selective recycle bioreactors.

    Ogden, K L; Davis, R H

    1991-02-20

    A new plasmid construct has been used in conjunction with selective recycle to successfully maintain otherwise unstable plasmid-bearing E. coli cells in a continuous bioreactor and to produce significant amounts of the plasmid-encoded protein beta-lactamase. The plasmid is constructed so that pilin expression, which leads to bacterial flocculation, is under control of the tac operon. The plasmid-bearing cells are induced to flocculate in the separator, whereas cell growth and product synthesis occur in the main fermentation vessel without the inhibiting effects of pilin production. Selective recycle allows for the maintenance of the plasmid-bearing cells by separating flocculent, plasmid-bearing cells from nonflocculent, segregant cells in an inclined settler, and recycling only the plasmid-bearing cells to the reactor. As a result, product expression levels are maintained that are more than ten times the level achieved without selective recycle. All experimental data agree well with theoretical predictions. PMID:18597374

  12. DISTRIBUTION OF PLASMIDS IN GROUNDWATER BACTERIA

    Bacteria isolated from groundwater aquifer core materials of pristine aquifers at Lula and Pickett, Oklahoma, and from a site with a history of aromatic hydrocarbon contamination and natural renovation located at Conroe, Texas, were screened for the presence of plasmid Deoxyribon...

  13. Plasmid mediated quinolone resistance in Enterobacteriaceae

    Veldman, K.T.

    2014-01-01

    This thesis describes the occurrence of Plasmid Mediated Quinolone Resistance (PMQR) in Salmonella and E. coli from The Netherlands and other European countries. Furthermore, the genetic background of these genes was characterized. Fluoroquinolones are widely used antibiotics in both human and veter

  14. Plasmid DNA entry into postmitotic nuclei of primary rat myotubes.

    Dowty, M E; Williams, P; Zhang, G.; Hagstrom, J E; Wolff, J A

    1995-01-01

    These studies were initiated to elucidate the mechanism of DNA nuclear transport in mammalian cells. Biotin- or gold-labeled plasmid and plasmid DNA expression vectors for Escherichia coli beta-galactosidase or firefly luciferase were microinjected into the cytoplasm of primary rat myotubes in culture. Plasmid DNA was expressed in up to 70% of the injected myotubes, which indicates that it entered intact, postmitotic nuclei. The nuclear transport of plasmid DNA occurred through the nuclear po...

  15. The influence of biofilms in the biology of plasmids

    Cook, Laura C.C.; Dunny, Gary M.

    2014-01-01

    The field of plasmid biology has historically focused on bacteria growing in liquid culture. Surface attached communities of bacterial biofilms have recently been understood to be the normal environment of bacteria in the natural world. Thus, studies examining plasmid replication, maintenance, and transfer in biofilms are essential for a true understanding of bacterial plasmid biology. This chapter reviews the current knowledge of the interplay between bacterial biofilms and plasmids, focusin...

  16. The 2 micron plasmid purloins the yeast cohesin complex

    Mehta, Shwetal; Yang, Xian Mei; Chan, Clarence S.; Dobson, Melanie J.; Jayaram, Makkuni; Velmurugan, Soundarapandian

    2002-01-01

    The yeast 2 micron plasmid achieves high fidelity segregation by coupling its partitioning pathway to that of the chromosomes. Mutations affecting distinct steps of chromosome segregation cause the plasmid to missegregate in tandem with the chromosomes. In the absence of the plasmid stability system, consisting of the Rep1 and Rep2 proteins and the STB DNA, plasmid and chromosome segregations are uncoupled. The Rep proteins, acting in concert, recruit the yeast cohesin complex to the STB locu...

  17. Linear and Circular Plasmid Content in Borrelia burgdorferi Clinical Isolates

    Iyer, Radha; Kalu, Ogori; Purser, Joye; Norris, Steven; Stevenson, Brian; Schwartz, Ira

    2003-01-01

    The genome of Borrelia burgdorferi, the etiologic agent of Lyme disease, is composed of a linear chromosome and more than 20 linear and circular plasmids. Typically, plasmid content analysis has been carried out by pulsed-field gel electrophoresis and confirmed by Southern hybridization. However, multiple plasmids of virtually identical sizes (e.g., lp28 and cp32) complicate the interpretation of such data. The present study was undertaken to investigate the complete plasmid complements of B....

  18. Postsegregational killing does not increase plasmid stability but acts to mediate the exclusion of competing plasmids

    Cooper, Tim F; Heinemann, Jack A.

    2000-01-01

    Postsegregational killing (PSK) systems consist of a tightly linked toxin–antitoxin pair. Antitoxin must be continually produced to prevent the longer lived toxin from killing the cell. PSK systems on plasmids are widely believed to benefit the plasmid by ensuring its stable vertical inheritance. However, experimental tests of this “stability” hypothesis were not consistent with its predictions. We suggest an alternative hypothesis to explain the evolution of PSK: ...

  19. Plasmid Capture by the Bacillus thuringiensis Conjugative Plasmid pXO16▿

    Timmery, Sophie; Modrie, Pauline; Minet, Olivier; Mahillon, Jacques

    2009-01-01

    Conjugation, mobilization, and retromobilization are three related mechanisms of horizontal gene transfer in bacteria. They have been extensively studied in gram-negative species, where retromobilization, the capture of DNA from a recipient by a donor cell, was shown to result from two successive steps: the transfer of the conjugative plasmid from the donor to the recipient followed by the retrotransfer of the mobilizable plasmid to the donor. This successive model was established for gram-ne...

  20. Compositional discordance between prokaryotic plasmids and host chromosomes

    van Kampen Antoine HC

    2006-02-01

    Full Text Available Abstract Background Most plasmids depend on the host replication machinery and possess partitioning genes. These properties confine plasmids to a limited range of hosts, yielding a close and presumably stable relationship between plasmid and host. Hence, it is anticipated that due to amelioration the dinucleotide composition of plasmids is similar to that of the genome of their hosts. However, plasmids are also thought to play a major role in horizontal gene transfer and thus are frequently exchanged between hosts, suggesting dinucleotide composition dissimilarity between plasmid and host genome. We compared the dinucleotide composition of a large collection of plasmids with that of their host genomes to shed more light on this enigma. Results The dinucleotide frequency, coined the genome signature, facilitates the identification of putative horizontally transferred DNA in complete genome sequences, since it was found to be typical for a certain genome, and similar between related species. By comparison of the genome signature of 230 plasmid sequences with that of the genome of each respective host, we found that in general the genome signature of plasmids is dissimilar from that of their host genome. Conclusion Our results show that the genome signature of plasmids does not resemble that of their host genome. This indicates either absence of amelioration or a less stable relationship between plasmids and their host. We propose an indiscriminate lifestyle for plasmids preserving the genome signature discordance between these episomes and host chromosomes.

  1. Conjugative plasmids: Vessels of the communal gene pool

    Norman, Anders; Hansen, Lars H.; Sørensen, Søren Johannes

    2009-01-01

    over large taxonomic distances. These plasmids are collections of discrete regions of genes that function as ‘backbone modules' to undertake different aspects of overall plasmid maintenance and propagation. Conjugative plasmids often carry suites of ‘accessory elements' that contribute adaptive traits...

  2. Competition between Plasmid-Bearing and Plasmid-Free Organisms in a Chemostat with Pulsed Input and Washout

    Sanling Yuan; Yu Zhao; Anfeng Xiao

    2009-01-01

    We consider a model of competition between plasmid-bearing and plasmid-free organisms in the chemostat with pulsed input and washout. We investigate the subsystem with nutrient and plasmid-free organism and study the stability of the boundary periodic solutions, which are the boundary periodic solutions of the system. The stability analysis of the boundary periodic solution yields the invasion threshold of the plasmid-bearing organism. By using the standard techniques of bifurcation theory, w...

  3. Co-segregation of yeast plasmid sisters under monopolin-directed mitosis suggests association of plasmid sisters with sister chromatids

    Liu, Yen-Ting; Ma, Chien-Hui; Jayaram, Makkuni

    2013-01-01

    The 2-micron plasmid, a high copy extrachromosomal element in Saccharomyces cerevisiae, propagates itself with nearly the same stability as the chromosomes of its host. Plasmid stability is conferred by a partitioning system consisting of the plasmid-coded proteins Rep1 and Rep2 and a cis-acting locus STB. Circumstantial evidence suggests that the partitioning system couples plasmid segregation to chromosome segregation during mitosis. However, the coupling mechanism has not been elucidated. ...

  4. The Residence Life Cinema.

    Dungan, Jane Fidler; Elion, Audrey; Gusmano, Phil

    1997-01-01

    Explores the implementation, results, and the limitations of the Residence Life Cinema program at the University of Memphis. Claims that such programs offer an innovative method for fostering student development by utilizing movies to stimulate affective and cognitive processes in students--processes that may not occur without a catalyst. (RJM)

  5. A comparison of the modern seismogenic Nankai mega-splay fault and the exhumed ancient mega-splay fault, the Nobeoka thrust

    Kimura, G.; Hamahashi, M.; Yamaguchi, A.; Saito, S.; Fukuchi, R.; Kameda, J.; Hamada, Y.; Fujimoto, K.; Hashimoto, Y.; Hina, S.; Eida, M.; Kitamura, Y.

    2012-12-01

    Mega-splay fault branched from plate boundary megathrust in subduction zone is located around the border between outer and inner wedges and is considered to cause great earthquake and tsunami such as 1960 Alaska earthquake, 1944 and 1946 Nankai-Tonankai earthquakes, and 2004 Sumatra earthquakes. Therefore, understanding the fault mechanics of the mega-splay fault is essential toward assessing their role in the plate boundary processes and seismo-tsunamigenesis. Seismic reflection studies for the mega-splay fault in 2D and 3D in the Nankai forearc present the reflector with negative or positive polarities with various amplitudes and suggest complicated petrophysical properties and condition of the fault and its surroundings. The Nankai mega-splay fault at a depth of ~5km is going to be drilled and cored by Integrated Ocean Drilling Program, NantroSEIZE experiments and is expected for great progress of understanding of the fault mechanics. Deep portion of the megasplay fault and its connection to the plate boundary megathrust is, however, impossible to be accessed by direct drilling. Far and near field geophysical observation is therefore only way to access the modern and active megasplay fault. On-land exhumed and fossilized mega-splay faults, on the other hand, give a clue for the fault mechanics when they were active in depth although the exhumation and fossilization process modifies their primary properties due to physico-chemical weathering and crack opening by unloading. Our previous studies from the Nobeoka thrust in Kyushu, southwest Japan present well-preservation of primary faulting processes and clear contrast of physical property between the hanging wall and footwall. We have conducted the seismic, drilling, coring and logging investigation into the Nobeoka thrust to the depth of ~250 m including ~40m hanging wall and ~210 m footwall. The coring was ~99% recovery and full logging was successful. The result of the logging together with triangular S

  6. THE INTEGRATED STATE OF THE ROLLING-CIRCLE PLASMID PTB913 IN THE COMPOSITE BACILLUS PLASMID PTB19

    OSKAM, L; HILLENGA, DJ; VENEMA, G; BRON, S

    1992-01-01

    pTB19, a 27 kb plasmid originating from a thermophilic Bacillus species, contains integrated copies of two rolling-circle type plasmids on a 10.6 kb DNA fragment. In the present study we analysed the part of pTB19 that contains the rolling-circle plasmid pTB913 and the region in between the two roll

  7. A Mega-project on Space Weather Monitoring Gets the Green Light

    2005-01-01

    @@ A 200-million-yuan ($25 million) mega-project of science research on space weather monitoring, namely the Meridian Space Weather Moni toting Project (Meridian Project for short), proposed by several research institutes and universities in China has been approved recently by the Chinese government.

  8. The bidding paradox: why rational politicians still want to bid for mega sports events

    de Nooij, M.; van den Berg, M.R.

    2013-01-01

    This paper discusses reasons why politicians still favor hosting mega events despite the discouraging evidence regarding their financial benefits: (1) early political enthusiasm, (2) tying side-projects to the bid to raise political support, (3) biased reading of history, (4) the winners curse, (5)

  9. Identify the costs and benefits of bidding for the hosting of a mega-event

    熊淑一

    2014-01-01

    A mega-event which attracts attention from all over the world might have both positive and negative effect on the hosting city (Syme et al.1989).The aim of this paper is to evaluate the costs and benefits of bidding for the hosting of mega-events.It identified legacies of bidding for the hosting of mega-events from following four aspects,which are economics,public life,infrastructure,and image of the hosting city,with the methods of literature review,logic analysis,examples,and contrast analysis.The result of this study unveiled that when a city bids for hosting a mega-event for the purpose of stimulating its local economic,improving civic pride,developing basic public infrastructures,and enhance its city image to the worldwide,it also needs to pay attention to potential drawbacks of hosting the event,such as debt crisis,interfering the daily life of the public.

  10. Of Mega-Malls and Soft-Shelled Turtles: Deep Ecological Education To Counter Homogeneity.

    Quinn, Michael S.; Scott, Jennifer R.

    1997-01-01

    Attempts to compare the modern retail/entertainment experience of the mega-mall to a singular encounter with a wild animal in its native community context. Explores the differences between seeking amusement and seeking happiness as well as the capacity for experiencing joy. Concludes with some implications of these experiences to the…

  11. Building the MEGA Image-Subtraction Pipeline in the Era of Virtual Observatories

    Alves, D R; Bergier, A; Cseresnjes, P; Gersch, A; Alves, David R.

    2003-01-01

    We are inventing techniques to increase variable source yields from image subtraction analyses. An example innovation that will improve detection efficiency by about 10% is presented. The MEGA image-subtraction pipeline is a software instrument that could evolve into a tool to do microlensing science with virtual observatories.

  12. Unemployment, Entrepreneurial Education and Mega Universities: Challenges to Expanding Access in Education in Nigeria University

    Undie, John Atewhoble; Okafor, Victor

    2014-01-01

    In fundamental economics, individuals acquired education for two broad reasons, as an investment and as consumption. The investment function of education has continued to create tension for job search leading to cases of unemployment. Entrepreneurship education and establishment of mega universities have been identified as panaceas. This paper…

  13. The economic performance of supply chain(s) served by the mega freight transport vehicles

    Janic, M.

    2014-01-01

    This paper deals with the economic performances of supply chain(s) served by different including the mega freight transport vehicles. These performances are considered as a dimension of the supply chain’s sustainability together with the infrastructural, technical/technological, operational, environ

  14. (Mega)structures: Structural Imagination and New Spatial Visions, 1945- 1970

    Pizza de Nanno, Antonio

    2012-01-01

    Reseña 2: Symposium. *(Mega)structures: Structural Imagination and New Spatial Visions, 1945- 1970: recorridos a partir del atlas mnemosyne. Seminario Internacional dirigido por Alessandro De Magistris, Federico Deambrosis y Antonio Pizza. Dipartimento di Architettura e Pianificazione Politecnico di Milano, 22-23 novembre 2011.

  15. Facilitators and Inhibitors of Bidding and Hosting Mega Sporting Events in Iran

    Mohammad Koosha

    2013-01-01

    Full Text Available Legacies of hosting mega sporting events have been attempting idea for both the developed and the developing nations to participate in "bidding" competitions. Through mega sporting events bid process, despite all defined essentials infrastructures, the viewpoint of local organizing committee play a significant role to propose a winning bid and delivery of successful games. This research aims to identify the facilitators and inhibitors of bidding and hosting mega sporting events in Iran from the viewpoints of the Iranian distinguished professors in the sport management and the sport executive managers. Researcher questionnaire and Semi-structure interviews were conducted to gain in depth data. The results indicated that Iran currently has limitations to have winning bid and successful host mega sporting events; even though, it possesses the potential to do so. Public interest and support for hosting the sport events, good security condition against terrorism and tourist attractions are known as the strong points and he challenges and inhibitors which Iran is faced with includes: the qualitative and quantitative weakness in the transportation infrastructures and sport venues, west countries economic boycott, unstable government support and the lack of long-run planning for the events.

  16. OER on the Asian Mega Universities: Developments, Motives, Openness, and Sustainability

    Farisi, Mohammad Imam

    2013-01-01

    The OER movement originated and integrated into ODE developments. Mega Universities (MUs) are among the most important of ODE providers worldwide should be to be the primary organizations for providing access to OER. So far, however, in-depth studies on OER developments in the Asian MUs were very limited. This study focuses on the developments,…

  17. Mega-event strategy as a tool of urban transformation: Sydney's experience

    Chen, Y.; Spaans, M.

    2009-01-01

    Mega-event strategies and their impact on host cities have drawn increasing interest, as organising large-scale urban events has become part of a deliberate urban policy strategy to promote local economic growth and put the host city on the world agenda. Thus far, the research addressing what strate

  18. Mega Events and Urban Conflicts in Valencia, Spain: Contesting the New Urban Modernity

    Luis del Romero Renau

    2011-01-01

    Full Text Available The local and regional administrations of Valencia, Spain, have been working to position the city, which is the country's third largest in terms of population, as an international destination for luxury tourism through sports mega-events. Valencia's new image as a “mega-events city” has been criticized by citizens and urban-based movements on social and environmental grounds, resulting in various urban conflicts. The aim of this paper is to examine the causes and dynamics of these conflicts in order to understand the discourse and rationales behind the criticism. This will also include an examination of the ways in which these conflicts were managed by the local administration, especially in the months leading up to and during the events, when the city was receiving worldwide media coverage. Researching local print media, we focused on urban conflicts that took place in Valencia between 1995 and 2010. We also looked at the performance of urban-based movements and the administration as conflicting actors. One of the main findings was the strong consensual authoritarianism manifested by the administration and mega-event organizers when making decisions and managing urban conflicts related to mega-events.

  19. (Mega)structures: Structural Imagination and New Spatial Visions, 1945- 1970

    Pizza de Nanno, Antonio

    2012-01-01

    Reseña 2: Symposium. *(Mega)structures: Structural Imagination and New Spatial Visions, 1945- 1970: recorridos a partir del atlas mnemosyne. Seminario Internacional dirigido por Alessandro De Magistris, Federico Deambrosis y Antonio Pizza. Dipartimento di Architettura e Pianificazione Politecnico di Milano, 22-23 novembre 2011. Peer Reviewed

  20. Stable inheritance of plasmid R1 requires two different loci.

    Gerdes, K; Larsen, J E; Molin, S

    1985-01-01

    The largest EcoRI fragment from plasmid R1 mediates a stability phenotype which is required to ensure the stable inheritance of this low-copy-number plasmid. When covalently linked to small, unstable R1 derivatives, this fragment makes the plasmids as stable as the wild-type R1 plasmid. A genetic analysis showed that two independently acting stabilization functions are encoded by this EcoRI fragment, both of which have the potential of partial stabilization of mini-R1 plasmids. The two loci a...

  1. DNA restriction-modification systems mediate plasmid maintenance.

    Kulakauskas, S; Lubys, A; Ehrlich, S. D.

    1995-01-01

    Two plasmid-carried restriction-modification (R-M) systems, EcoRI (from pMB1 of Escherichia coli) and Bsp6I (from pXH13 of Bacillus sp. strain RFL6), enhance plasmid segregational stability in E. coli and Bacillus subtilis, respectively. Inactivation of the endonuclease or the presence of the methylase in trans abolish the stabilizing activity of the R-M systems. We propose that R-M systems mediate plasmid segregational stability by postsegregational killing of plasmid-free cells. Plasmid-enc...

  2. Non-climatic signal in ice core records: lessons from Antarctic mega-dunes

    Ekaykin, A.; Eberlein, L.; Lipenkov, V.; Popov, S.; Scheinert, M.; Schröder, L.; Turkeev, A.

    2015-12-01

    We present the results of glaciological investigations in the mega-dune area located 30 km to the east from Vostok Station (central East Antarctica) implemented during the 58th, 59th and 60th Russian Antarctic Expedition (January 2013-January 2015). Snow accumulation rate and isotope content (δD, δ18O and δ17O) were measured along the 2 km profile across the mega-dune ridge accompanied by precise GPS altitude measurements and GPR survey. It is shown that the spatial variability of snow accumulation and isotope content covaries with the surface slope. The accumulation rate regularly changes by one order of magnitude within the distance < 1 km, with the reduced accumulation at the leeward slope of the dune and increased accumulation in the hollow between the dunes. At the same time, the accumulation rate averaged over the length of a dune wave (22 mm we) corresponds well with the value obtained at Vostok Station, which suggests no additional wind-driven snow sublimation in the mega-dunes compared to the surrounding plateau. The snow isotopic composition is in negative correlation with the snow accumulation. Analyzing dxs/δD and 17O-excess/δD slopes, we conclude that the spatial variability of the snow isotopic composition in the mega-dune area could be explained by post-depositional snow modifications. Using the GPR data, we estimated the apparent dune drift velocity (4.6 ± 1.1 m yr-1). The full cycle of the dune drift is thus about 410 years. Since the spatial anomalies of snow accumulation and isotopic composition are supposed to drift with the dune, an ice core drilled in the mega-dune area would exhibit the non-climatic 410 year cycle of these two parameters. We simulated a vertical profile of snow isotopic composition with such a non-climatic variability, using the data on the dune size and velocity. This artificial profile is then compared with the real vertical profile of snow isotopic composition obtained from a core drilled in the mega-dune area. We

  3. Radar scattering of linear dunes and mega-yardangs: Application to Titan

    Paillou, Philippe; Seignovert, Benoît; Radebaugh, Jani; Wall, Stephen

    2016-05-01

    The Ku-band (13.8 GHz - 2.2 cm) RADAR instrument onboard the Cassini-Huygens spacecraft has revealed the richness of the surface of Titan, as numerous seas, lakes, rivers, cryo-volcanic flows and vast dune fields have been discovered. Linear dunes are a major geomorphological feature present on Titan, covering up to 17% of its surface, mainly in equatorial regions. However, the resolution of the RADAR instrument is not good enough to allow a detailed study of the morphology of these features. In addition, other linear wind-related landforms, such as mega-yardangs (linear wind-abraded ridges formed in cohesive rocks), are likely to present a comparable radar signature that could be confused with the one of dunes. We conducted a comparative study of the radar radiometry of both linear dunes and mega-yardangs, based on representative terrestrial analogues: the linear dunes located in the Great Sand Sea in western Egypt and in the Namib Desert in Namibia, and the mega-yardangs observed in the Lut Desert in eastern Iran and in the Borkou Desert in northern Chad. We analysed the radar scattering of both terrestrial linear dunes and mega-yardangs, using high-resolution radar images acquired by the X-band (9.6 GHz - 3.1 cm) sensor of the TerraSAR-X satellite. Variations seen in the radar response of dunes are the result of a contrast between the dune and interdune scattering, while for mega-yardangs these variations are the result of a contrast between ridges and erosion valleys. We tested a simple surface scattering model, with parameters derived from the local topography and surface roughness estimates, to accurately reproduce the radar signal variations for both landforms. It appears that we can discriminate between two types of dunes - bare interdunes as in Egypt and sand-covered interdunes as in Namibia, and between two types of mega-yardangs - young yardangs as in Iran and older ones as in Chad. We applied our understanding of the radar scattering to the analysis of

  4. Role of Plasmid in Production of Acetobacter Xylinum Biofilms

    Abbas Rezaee

    2005-01-01

    Full Text Available Acetobacter xylinum has the ability to produce cellulotic biofilms. Bacterial cellulose is expected to be used in many industrial or biomedical materials for its unique characteristics. A. xylinum contains a complex system of plasmid DNA molecules. A 44 kilobases (kb plasmid was isolated in wild type of A. xylinum. To improve the cellulose producing ability of A. xylinum, role of the plasmid in production of cellulose was studied. The comparisons between wild type and cured cells of A. xylinum showed that there is considerably difference in cellulose production. In order to study the relationship between plasmid and the rate of cellulose production, bacteria were screened for plasmid profile by a modified method for preparation of plasmid. This method yields high levels of pure plasmid DNA that can be used for common molecular techniques, such as digestion and transformation, with high efficiency.

  5. Interdecadal variability of the mega-ENSO-NAO synchronization in winter

    Wu, Zhiwei; Zhang, Peng

    2015-08-01

    Mega-El Niño-Southern Oscillation (ENSO) and the North Atlantic Oscillation (NAO), as two principal components of the global air-sea coupling system, may have synchronous or out-synchronous fluctuations during different epochs. Understanding such connection change is instrumental for climate prediction, particularly the decadal prediction. Results in this study show that mega-ENSO has experienced a notable inter-decadal change in its linkage with the winter NAO during the past 56 years: mega-ENSO was significantly correlated with the NAO during 1957-1981 (or synchronous epoch), while such correlation has broken down since 1982 (or out-synchronous epoch). This marked change might be attributed to a sea surface temperature (SST) forcing change in the North Atlantic, based on the observational and numerical evidences in this study. The synchronous epoch is concurrent with the anomalous tropical North Atlantic (TNA) SST forcing, whereas the out-synchronous epoch is associated with the anomalous extra-tropical North Atlantic (XNA) SST forcing. Two possible reasons may explain how the synchronous behaviors between mega-ENSO and the NAO were tied to the TNA SST anomaly (SSTA). There is a positive feedback between the TNA SSTA and the NAO-like atmosphere anomalies, which helps to "prolong" the NAO impacts from the developing phase through mature phase of mega-ENSO. Additionally, the TNA SSTA itself may induce a NAO-like atmosphere anomaly. Since 1982, the TNA SSTA has been replaced by the XNA SSTA and the latter primarily favors a NAO-neutral state in the atmosphere, which ends the synchronous epoch.

  6. Deficient sumoylation of yeast 2-micron plasmid proteins Rep1 and Rep2 associated with their loss from the plasmid-partitioning locus and impaired plasmid inheritance.

    Jordan B Pinder

    Full Text Available The 2-micron plasmid of the budding yeast Saccharomyces cerevisiae encodes copy-number amplification and partitioning systems that enable the plasmid to persist despite conferring no advantage to its host. Plasmid partitioning requires interaction of the plasmid Rep1 and Rep2 proteins with each other and with the plasmid-partitioning locus STB. Here we demonstrate that Rep1 stability is reduced in the absence of Rep2, and that both Rep proteins are sumoylated. Lysine-to-arginine substitutions in Rep1 and Rep2 that inhibited their sumoylation perturbed plasmid inheritance without affecting Rep protein stability or two-hybrid interaction between Rep1 and Rep2. One-hybrid and chromatin immunoprecipitation assays revealed that Rep1 was required for efficient retention of Rep2 at STB and that sumoylation-deficient mutants of Rep1 and Rep2 were impaired for association with STB. The normal co-localization of both Rep proteins with the punctate nuclear plasmid foci was also lost when Rep1 was sumoylation-deficient. The correlation of Rep protein sumoylation status with plasmid-partitioning locus association suggests a theme common to eukaryotic chromosome segregation proteins, sumoylated forms of which are found enriched at centromeres, and between the yeast 2-micron plasmid and viral episomes that depend on sumoylation of their maintenance proteins for persistence in their hosts.

  7. Modeling sRNA-regulated Plasmid Maintenance

    Gong, Chen Chris

    2016-01-01

    We study a theoretical model for the toxin-antitoxin (hok/sok) mechanism for plasmid maintenance in bacteria. Toxin-antitoxin systems enforce the maintenance of a plasmid through post-segregational killing of cells that have lost the plasmid. Key to their function is the tight regulation of expression of a protein toxin by an sRNA antitoxin. Here, we focus on the nonlinear nature of the regulatory circuit dynamics of the toxin-antitoxin mechanism. The mechanism relies on a transient increase in protein concentration rather than on the steady state of the genetic circuit. Through a systematic analysis of the parameter dependence of this transient increase, we confirm some known design features of this system and identify new ones: for an efficient toxin-antitoxin mechanism, the synthesis rate of the toxin's mRNA template should be lower that of the sRNA antitoxin, the mRNA template should be more stable than the sRNA antitoxin, and the mRNA-sRNA complex should be more stable than the sRNA antitoxin. Moreover, ...

  8. Mega starbirth cluster is biggest, brightest and hottest ever seen

    2003-10-01

    -European Coordinating Facility in Germany, and a team of international co-authors report the discovery in the 20 October 2003 issue of the Astrophysical Journal. The mega-cluster of stars appears as a puzzling red arc behind a distant galaxy cluster 5400 million light-years away in the northern constellation of Lynx. The arc is the stretched and magnified image of a mysterious celestial object about 12 000 million light-years away (at a redshift of 3.36), far beyond the cluster of galaxies. This means that the remote source existed when the Universe was less than 2000 million years old. Fosbury and colleagues first tried to identify the arc by analysing the light from the object, but the team was not able to recognise the pattern of colours in the spectral signature of the remote object. While looking for matches with the colour spectrum, Fosbury realised that the light was related to that of the nearby Orion Nebula, a star-forming region in our own Milky Way. However where the Orion Nebula is powered by only four hot and bright blue stars, the Lynx Arc must contain around a million such stars! Furthermore, the spectrum shows that the stars in the Lynx Arc are more than twice as hot as the Orion Nebula's central stars, with surface temperatures up to 80 000°C. Though there are much bigger and brighter star-forming regions than the Orion Nebula in our local Universe, none are as bright as the Lynx Arc, nor do they contain such large numbers of hot stars. Even the most massive, normal nearby stars are no hotter than around 40 000°C. However, stars forming from the original, pristine gas in the early Universe can be more massive and consequently much hotter - perhaps up to 120 000°C. The earliest stars may have been as much as several hundred solar masses, but the chemical make-up of the Universe today prevents stars from forming beyond about 100 solar masses. Such 'primordial' super-hot stars are thought to be the first luminous objects to condense after the Big Bang cooled

  9. Lawful Permanent Residents - Annual Report

    Department of Homeland Security — A lawful permanent resident (LPR) or 'green card' recipient is defined by immigration law as a person who has been granted lawful permanent residence in the United...

  10. Determination of Plasmid Segregational Stability in a Growing Bacterial Population.

    Kramer, M Gabriela

    2016-01-01

    Bacterial plasmids are extensively used as cloning vectors for a number of genes for academic and commercial purposes. Moreover, attenuated bacteria carrying recombinant plasmids expressing genes with anti-tumor activity have shown promising therapeutic results in animal models of cancer. Equitable plasmid distribution between daughter cells during cell division, i.e., plasmid segregational stability, depends on many factors, including the plasmid copy number, its replication mechanism, the levels of recombinant gene expression, the type of bacterial host, and the metabolic burden associated with all these factors. Plasmid vectors usually code for antibiotic-resistant functions, and, in order to enrich the culture with bacteria containing plasmids, antibiotic selective pressure is commonly used to eliminate plasmid-free segregants from the growing population. However, administration of antibiotics can be inconvenient for many industrial and therapeutic applications. Extensive ongoing research is being carried out to develop stably-inherited plasmid vectors. Here, I present an easy and precise method for determining the kinetics of plasmid loss or maintenance for every ten generations of bacterial growth in culture. PMID:26846807

  11. Plasmid transfer between bacteria in soil microcosms and the field

    Eric Smit

    1997-01-01

    Full Text Available In ibis review factors influencing conjugal plasmid transfer between bacteria and the possible role of naturally occurring selftransmissible plasmide for the dissemination of recombinant DNA in soil will be discussed. In microcosm studies, plasmid transfer between various species of introduced bacteria has been detected. Moreover, plamid transfer to indigenous soil micoorganisms was observed. Soil is an oligotrophic environment and plasmid transfer occurred mainly under conditions which were nutritionally favourable for bacteria, such as in the plant rhizosphere and in the presence of clay minerais or added nutrients. Mobilizable plasmids, lacking the ability to transfer themselves, have been reported to be transferred in the presence of selftransmissible plasmids. A study comparing conjugal transfer in microcosme with those in the field revealed that the transfer rates found in microcosme and in the field were similar. Transfer of chromosomal DNA by plasmid RP4 could only be shown on filters and was not observed in soil. Transfer of plasmids carrying biodegradative genes appeared to be favoured in the presence of the compound that can be degraded. Evidence was found for the presence of naturally-occurring selftransmissible plasmids in bacteria in the rhizosphere which could mobilize recombinant plasmids.

  12. Characterization of toxin plasmids in Clostridium perfringens type C isolates.

    Gurjar, Abhijit; Li, Jihong; McClane, Bruce A

    2010-11-01

    Clostridium perfringens type C isolates cause enteritis necroticans in humans or necrotizing enteritis and enterotoxemia in domestic animals. Type C isolates always produce alpha toxin and beta toxin but often produce additional toxins, e.g., beta2 toxin or enterotoxin. Since plasmid carriage of toxin-encoding genes has not been systematically investigated for type C isolates, the current study used Southern blot hybridization of pulsed-field gels to test whether several toxin genes are plasmid borne among a collection of type C isolates. Those analyses revealed that the surveyed type C isolates carry their beta toxin-encoding gene (cpb) on plasmids ranging in size from ∼65 to ∼110 kb. When present in these type C isolates, the beta2 toxin gene localized to plasmids distinct from the cpb plasmid. However, some enterotoxin-positive type C isolates appeared to carry their enterotoxin-encoding cpe gene on a cpb plasmid. The tpeL gene encoding the large clostridial cytotoxin was localized to the cpb plasmids of some cpe-negative type C isolates. The cpb plasmids in most surveyed isolates were found to carry both IS1151 sequences and the tcp genes, which can mediate conjugative C. perfringens plasmid transfer. A dcm gene, which is often present near C. perfringens plasmid-borne toxin genes, was identified upstream of the cpb gene in many type C isolates. Overlapping PCR analyses suggested that the toxin-encoding plasmids of the surveyed type C isolates differ from the cpe plasmids of type A isolates. These findings provide new insight into plasmids of proven or potential importance for type C virulence. PMID:20823204

  13. Challenges to publishing pharmacy resident research projects from the perspectives of residency program directors and residents

    Olson KL

    2013-09-01

    Full Text Available Objective: To identify barriers to completing and publishing pharmacy residency research projects from the perspective of program directors and former residents.Methods: This was a cross-sectional survey of pharmacy residency program directors and former post-graduate year one and two residents. Directors of pharmacy residency programs whose residents present their projects at the Western States Conference (n=216 were invited to complete an online survey and asked to forward the survey to former residents of their program in 2009, 2010, or 2011. The survey focused on four broad areas: 1 demographic characteristics of the residency programs, directors, and residents; 2 perceived value of the research project; 3 perceived barriers with various stages of research; and 4 self-identified barriers to successful research project completion and publication.Results: A total of 32 program directors and 98 residents completed the survey. The minority of programs offered formal residency research training. Both groups reported value in the research project as part of residency training. Significantly more directors reported obtaining institutional review board approval and working through the publication process as barriers to the research project (46.7% vs. 22.6% and 73.3% vs. 43.0%, respectively p<0.05 while residents were more likely to report collecting and analyzing the data as barriers (34.4% vs. 13.3% and 39.8% vs. 20.0%, respectively, p<0.05. Both groups self-identified time constraints and limitations in study design or quality of the study as barriers. However, while program directors also indicated lack of resident motivation (65.5%, residents reported lack of mentorship or program structural issues (43.3%. Conclusion: Overall, while both groups found value in the residency research projects, there were barriers identified by both groups. The results of this study may provide areas of opportunity for improving the quality and publication rates of

  14. Design of Hollow Reinforced Concrete Columns in the Tubed Mega Frame : Dimensionering av ihåliga armerade betongpelare i ”Tubed Mega Frame”

    Tönseth, David; Welchermill, Kristian

    2014-01-01

    A new concept for the structural system for tall buildings, called the “Tubed Mega Frame”, has been developed by Tyréns AB. The structure consists of several hollow reinforced concrete columns at the perimeter of the building and at certain levels, the columns are tied together with perimeter walls. Together they carry all the vertical and lateral loads. A purpose of the new concept is to eliminate the core in the center of the building which allows utilizing more floor spacing compared with...

  15. Evolving seismogenic plate boundary megathrust and mega-splay faults in subduction zone (Invited)

    Kimura, G.; Hamahashi, M.; Fukuchi, R.; Yamaguchi, A.; Kameda, J.; Kitamura, Y.; Hashimoto, Y.; Hamada, Y.; Saito, S.; Kawasaki, R.

    2013-12-01

    Understanding the fault mechanism and its relationship to the sesimo-tsunamigenesis is a key of the scientific targets of subduction zone and therefore NantroSEIZE project of IODP and future new drilling project of International Ocean Discovery Program keeps focusing on that. Mega-splay fault branched from plate boundary megathrust in subduction zone is located around the border between outer and inner wedges and is considered to cause great earthquake and tsunami such as 1960 Alaska earthquake, 1944 and 1946 Nankai-Tonankai earthquakes, and 2004 Sumatra earthquakes. Seismic reflection studies for the mega-splay fault in 2D and 3D in the Nankai forearc present the reflector with negative or positive polarities with various amplitudes and suggest complicated petrophysical properties and condition of the fault and its surroundings. The Nankai mega-splay fault at a depth of ~5km is going to be drilled and cored by NantroSEIZE experiments and is expected for great progress of understanding of the fault mechanics. Before drilling the really targeted seismogenic fault, we are conducting many exercises of geophysical and geological observations. The core-log-seismic integrated exercise for the exhumed mega-splay fault by drilling was operated for the Nobeoka thrust in the Shimanto Belt, Kyushu, Japan. The Nobeoka thrust was once buried in the depth >~10km and suffered maximum temperature >~300 dgree C. As the core recovery is ~99%, perfect correlation between the core and logging data is possible. Thickness of the fault zone is >200 m with a ~50 cm thick central fault core dividing the phyllitic hanging wall and the footwall of broken-melange like cataclasite. A-few-meter-thick discrete damage zones with fault cores are recognized by difference in physical properties and visual deformation textures at several horizons in the fault zone. Host rocks for those damaged zones are completely lithified cataclasites with abundant mineral veins, which record the older and deeper

  16. pB264, a small, mobilizable, temperature sensitive plasmid from Rhodococcus

    Currie Devin H

    2004-04-01

    Full Text Available Abstract Background Gram-positive bacteria of the genus Rhodococcus have shown an extraordinary capacity for metabolizing recalcitrant organic compounds. One hindrance to the full exploitation of Rhodococcus is the dearth of genetic tools available for strain manipulation. To address this issue, we sought to develop a plasmid-based system for genetic manipulation of a variety of Rhodococcus strains. Results We isolated and sequenced pB264, a 4,970 bp cryptic plasmid from Rhodococcus sp. B264-1 with features of a theta-type replication mechanism. pB264 was nearly identical to pKA22, a previously sequenced but uncharacterized cryptic plasmid. Derivatives of pB264 replicate in a diverse range of Rhodococcus species, showing that this plasmid does not bear the same host range restrictions that have been exhibited by other theta replicating plasmids. Replication or maintenance of pB264 is inhibited at 37°C, making pB264 useful as a suicide vector for genetic manipulation of Rhodococcus. A series of deletions revealed that ca. 1.3 kb from pB264 was sufficient to support replication and stable inheritance of the plasmid. This region includes two open reading frames that encode functions (RepAB that can support replication of pB264 derivatives in trans. Rhodococcus sp. B264-1 will mobilize pB264 into other Rhodococcus species via conjugation, making it possible to genetically modify bacterial strains that are otherwise difficult to transform. The cis-acting element (oriT required for conjugal transfer of pB264 resides within a ca. 0.7 kb region that is distinct from the regions responsible for replication. Conclusion Shuttle vectors derived from pB264 will be useful for genetic studies and strain improvement in Rhodococcus, and will also be useful for studying the processes of theta replication and conjugal transfer among actinomycetes.

  17. Unique type of plasmid maintenance function: postsegregational killing of plasmid-free cells.

    Gerdes, K; Rasmussen, P. B.; Molin, S

    1986-01-01

    The stability locus parB+ of plasmid R1 has been found to specify a unique type of plasmid maintenance function. Two genes, hok (host killing) and sok (suppressor of killing), are required for the stabilizing activity. The hok gene encodes a highly toxic gene product, whose overexpression causes a rapid killing and a concomitant dramatic change in morphology of the host cell. The other gene, sok, was found to encode a product that counteracts the hok gene-mediated killing. The parB+ region wa...

  18. Molecular characterization of "plasmid-free" antibiotic-resistant Haemophilus influenzae.

    Roberts, M C; Smith, A. L.

    1980-01-01

    We examined 14 multiresistant and 8 ampicillin- or tetracycline-resistant Haemophilus influenzae isolates and 4 ampicillin-resistant H. parainfluenzae isolates for plasmid deoxyribonucleic acid. Sixteen strains carried plasmids. Both "plasmid-free" and plasmid-carrying isolates transferred the antibiotic resistance by conjugation. All transconjugants carried plasmid deoxyribonucleic acid, suggesting that the apparent plasmid-free strains contained R plasmids encoding for antibiotic resistance.

  19. Resident Peritoneal NK cells

    Gonzaga, Rosemary; Matzinger, Polly; Perez-Diez, Ainhoa

    2011-01-01

    Here we describe a new population of NK cells that reside in the normal, un-inflamed peritoneal cavity. Phenotypically, they share some similarities with the small population of CD49b negative, CD27 positive immature splenic NK cells, and liver NK cells but differ in their expression of CD62L, TRAIL and EOMES. Functionally, the peritoneal NK cells resemble the immature splenic NK cells in their production of IFN-γ, GM-CSF and TNF-α and in the killing of YAC-1 target cells. We also found that ...

  20. MegaFlex Scale-Up Cost & Risk Reduction for >50kW Future Power Demands Project

    National Aeronautics and Space Administration — As the MegaFlex solar array is scaled for power demands greater than 50kW over the next 20 years and deployed load requirements remain high or increase, advanced...

  1. Parallels in portraits of leadership in mega churches of Gauteng (RSA) and Florida (USA) / Tsietsi John Maloma

    Maloma, Tsietsi John

    2011-01-01

    This thesis, Parallels in portraits of leadership in mega churches of Gauteng (RSA) and Florida (USA), the researcher, studied literature on church leadership with special reference to the leadership of mega churches. He researched issues relating to the nature of church leadership by exploring relevant Bible passages, the lives of certain Bible characters that the Bible presents as good but not perfect leaders, as well as relevant key theological conceptions of leadership. The study accordin...

  2. Pengaruh Inovasi Dan Jiwa Kewirausahaan Terhadap Keberhasilan Usaha (Studi Kasus: Pelaku UMKM Kuliner Chinatown Asia Mega Mas Medan)

    Mawaddah, Nadia Inda

    2016-01-01

    This research was conducted to examine the effect of innovation and entrepreneurs Spirit to business success (Case Study: SMEs Of Kuliner Chinatown Asia Mega Mas Medan). The research was conducted on SMEs in the Asia Mega Mas with a total sample of 35 businesses. The sampling method used in this research was saturated sampling by using descriptive analysis method, statistical analysis method whis consist of double linear regression analysis, simultaneous significance test (F-test), partial si...

  3. Defining a quantitative framework for evaluation and optimisation of the environmental impacts of mega-event projects

    Parkes, O.; Lettieri, P.; Bogle, I. D.

    2016-01-01

    This paper presents a novel quantitative methodology for the evaluation and optimisation of the environmental impacts of the whole life cycle of a mega-event project: construction and staging the event and post-event site redevelopment and operation. Within the proposed framework, a mathematical model has been developed that takes into account greenhouse gas (GHG) emissions resulting from use of transportation fuel, energy, water and construction materials used at all stages of the mega-event...

  4. Streamlined Purification of Plasmid DNA From Prokaryotic Cultures

    Pueschel, Laura; Li, Hongshan; Hymes, Matthew

    2011-01-01

    We describe the complete process of AcroPrep Advance Filter Plates for 96 plasmid preparations, starting from prokaryotic culture and ending with high purity DNA. Based on multi-well filtration for bacterial lysate clearance and DNA purification, this method creates a streamlined process for plasmid preparation. Filter plates containing silica-based media can easily be processed by vacuum filtration or centrifuge to yield appreciable quantities of plasmid DNA. Quantitative analyses determine ...

  5. Safety and efficacy of DNA vaccines: Plasmids vs. minicircles

    Stenler, Sofia; Blomberg, Pontus; Smith, Ci Edvard

    2014-01-01

    While DNA vaccination using plasmid vectors is highly attractive, there is a need for further vector optimization regarding safety, stability, and efficiency. In this commentary, we review the minicircle vector (MC), which is an entity devoid of plasmid bacterial sequences, as an alternative to the traditional plasmid construct. The commentary highlights the recent discovery by Stenler et al. (2014) that the small size of an MC enables improved resistance to the shearing forces associated wit...

  6. Pathogenomics of the Virulence Plasmids of Escherichia coli

    Johnson, Timothy J.; Lisa K. Nolan

    2009-01-01

    Summary: Bacterial plasmids are self-replicating, extrachromosomal elements that are key agents of change in microbial populations. They promote the dissemination of a variety of traits, including virulence, enhanced fitness, resistance to antimicrobial agents, and metabolism of rare substances. Escherichia coli, perhaps the most studied of microorganisms, has been found to possess a variety of plasmid types. Included among these are plasmids associated with virulence. Several types of E. col...

  7. Transformation of Bacillus thuringiensis protoplasts by plasmid deoxyribonucleic acid.

    Martin, P A; Lohr, J. R.; Dean, D H

    1981-01-01

    A method has been developed to transform plasmid deoxyribonucleic acid into protoplasts of the insect pathogen Bacillus thuringiensis. Protoplasts were formed by treatment of cells with lysozyme. The efficiency of formation of protoplasts was affected by the strain, the media, and the cell density. Deoxyribonucleic acid uptake was induced by polyethylene glycol. Deoxyribonucleic acid from the Staphylococcus aureus plasmid pC194 was used for transformation. Although this plasmid could not be i...

  8. Identification of plasmid partition function in coryneform bacteria.

    Kurusu, Y; Satoh, Y.; Inui, M.; Kohama, K; Kobayashi, M.; Terasawa, M.; Yukawa, H

    1991-01-01

    We have identified and characterized a partition function that is required for stable maintenance of plasmids in the coryneform bacteria Brevibacterium flavum MJ233 and Corynebacterium glutamicum ATCC 31831. This function is localized to a HindIII-NspV fragment (673 bp) adjacent to the replication region of the plasmid, named pBY503, from Brevibacterium stationis IFO 12144. The function was independent of copy number control and was not associated directly with plasmid replication functions. ...

  9. Comparative genetic organization of incompatibility group P degradative plasmids.

    Burlage, R S; Bemis, L A; Layton, A C; Sayler, G. S.; Larimer, F

    1990-01-01

    Plasmids that encode genes for the degradation of recalcitrant compounds are often examined only for characteristics of the degradative pathways and ignore regions that are necessary for plasmid replication, incompatibility, and conjugation. If these characteristics were known, then the mobility of the catabolic genes between species could be predicted and different catabolic pathways might be combined to alter substrate range. Two catabolic plasmids, pSS50 and pSS60, isolated from chlorobiph...

  10. Identify the costs and benefits of bidding for the hosting of a mega-event

    熊淑一

    2014-01-01

    A mega-event which attracts attention from all over the world might have both positive and negative effect on the hosting city(Syme et al.1989).The aim of this paper is to evaluate the costs and benefits of bidding for the hosting of megaevents.It identified legacies of bidding for the hosting of megaevents from following four aspects,which are economics,public life,infrastructure,and image of the hosting city,with the methods of literature review,logic analysis,examples,and contrast analysis.The result of this study unveiled that when a city bids for hosting a mega-event for the purpose of stimulating its local economic,improving civic pride,developing basic public infrastructures,and enhance its city image to the worldwide,it also needs to pay attention to potential drawbacks of hosting the event,such as debt crisis,interfering the daily life of the public.

  11. Nuclear background effects on plasma diagnostics for mega-joule class laser facility

    Estimating the vulnerability is a key challenge for plasma diagnostics designed to operate in radiative background associated with mega-joule class laser facilities. Since DT shots at OMEGA laser facility reproduce the perturbing source expected during the first 100 nanoseconds of a typical DT shot realized at National Ignition Facility (NIF) and Laser MegaJoule facility (LMJ), vulnerability of diagnostic elements such as optical relays or optical analyzers were experimentally studied and, if necessary, hardening approaches have been initiated to authorize their use at higher radiative constraints. Other facilities such as nuclear reactor or accelerator have been also used to estimate vulnerability issues as radiation induced emission of glasses or damage in multilayer coatings. (authors)

  12. The Moses, mega-Moses, and Armstrong illusions: integrating language comprehension and semantic memory.

    Shafto, M; MacKay, D G

    2000-09-01

    This study develops a new theory of the Moses illusion, observed in responses to general knowledge questions such as, "How many animals of each kind did Moses take on the Ark?" People often respond "two" rather than "zero" despite knowing that Noah, not Moses, launched the Ark. Our theory predicted two additional types of conceptual error demonstrated here: the Armstrong and mega-Moses illusions. The Armstrong illusion involved questions resembling, "What was the famous line uttered by Louis Armstrong when he first set foot on the moon?" People usually comprehend such questions as valid, despite knowing that Louis Armstrong was a jazz musician who never visited the moon. This Armstrong illusion was not due to misperceiving the critical words (Louis Armstrong), and occurred as frequently as the Moses illusion (with critical words embedded in identical sentential contexts), but less frequently than the mega-Moses illusion caused when Moses and Armstrong factors were combined. PMID:11228907

  13. Tunable mega-ampere electron current propagation in solids by dynamic control of lattice melt.

    MacLellan, D.A.; Carroll, D.C.; Gray, R J; Booth, N.; Burza, Matthias; Desjarlais, M. P.; Du, F.; Neely, D; Powell, H. W.; Robinson, A. P. L.; Scott, G. G.; Yuan, X.H.; Wahlström, Claes-Göran; McKenna, P

    2014-01-01

    The influence of lattice-melt-induced resistivity gradients on the transport of mega-ampere currents of fast electrons in solids is investigated numerically and experimentally using laser-accelerated protons to induce isochoric heating. Tailoring the heating profile enables the resistive magnetic fields which strongly influence the current propagation to be manipulated. This tunable laser-driven process enables important fast electron beam properties, including the beam divergence, profile, a...

  14. Multi-dimensional project management models for mega-construction companies and their applications

    2011-01-01

    China is undertaking an increasing number of mega-infrastructure projects along with its rapid economic expansion.For participants in these projects,understand how to adapt the proper and efficient management strategies is critical.In this paper,we propose three distinct models for companies under various background and management requirements,based on sophisticated data analysis and model development.Moreover,we successfully committed these models with hydropower development projects and achieved a great progress in enhancing management efficiency.

  15. Mitigation technique for use of CMOS image sensors in mega-joule class laser radiative environment

    Presented is a new mitigation technique to improve the radiation tolerance of CMOS image sensors to the radiation constraints associated to the fusion by inertial confinement experiments at mega-joule class laser facilities. Using the global reset mode, results acquired at the OMEGA facility show the efficiency of this technique to reduce by more than 70% the number of white pixels induced by the mixed 14 MeV neutron and gamma-ray pulse. (authors)

  16. Improving Customer Satisfaction and Customer Service Levels : Case: Nakumatt Mega City Supermarket

    Onyango, Shalotte Salmi

    2011-01-01

    The purpose of this thesis was to analyze the current customer satisfaction and service levels of the Nakumatt mega city Supermarket through an efficient customer satisfaction survey. It is important for a company to work towards satisfying customers’ needs and expectations in order to flourish. The main objective of the study was to discover solutions to improve customer satisfaction and to increase the level of Nakumatt customer services in order to retain and attract new customers. Ano...

  17. Computer Arithmetic Algorithms for Mega-Digit Floating Point Numbers' Precision

    Musbah J. Aqel; Mohammed H. Saleh

    2007-01-01

    IEEE standard 754 floating point is the most common representation used for floating point numbers, and many computer arithmetic algorithms are developed for basic operations on this standard. In this study, new computer algorithms are proposed to increase the precision range and to solve some problems that are available while using these algorithms. However, these algorithms provide an optional range of required accuracy (Mega-Digit precision) to meet new computer's applications.

  18. OER ON THE ASIAN MEGA UNIVERSITIES: DEVELOPMENTS, MOTIVES, OPENNESS, AND SUSTAINABILITY

    Mohammad Imam FARISI,

    2013-01-01

    The OER movement originated and integrated into ODE developments. Mega Universities (MUs) are among the most important of ODE providers worldwide should be to be the primary organizations for providing access to OER. So far, however, in-depth studies on OER developments in the Asian MUs were very limited. This study focuses on the developments, motives, openness, and sustainability of OER at the six MUs official websites in Asia. Data were collected for six months using the documentation tech...

  19. Sport mega-events: can legacies and development be equitable and sustainable?

    Jay Coakley; Doralice Lange de Souza

    2013-01-01

    Sport mega-events (SMEs) involve struggles to determine the definition of legacy and the outcome priorities that guide legacy planning, funding, and implementation processes. History shows that legacies reflect the interests of capital, and legacy benefits are enjoyed primarily, if not exclusively, by powerful business interests, a few political leaders, and organizations that govern high performance sports. This paper addresses challenges faced by cities and countries that host SMEs, and sho...

  20. Characterization of Chlamydia trachomatis Plasmid-Encoded Open Reading Frames

    Gong, Siqi; Yang, Zhangsheng; Lei, Lei; Shen, Li; Zhong, Guangming

    2013-01-01

    The recent success in transformation of Chlamydia trachomatis represents a major advancement in Chlamydia research. Plasmid-free C. trachomatis serovar L2 organisms can be transformed with chlamydial plasmid-based shuttle vectors pGFP::SW2 and pBRCT. Deletion of plasmid genes coding for Pgp1 to Pgp8 in pBRCT led to the identification of Pgp1, -2, -6, and -8 as plasmid maintenance factors; Pgp4 as a transcriptional regulator of chlamydial virulence-associated gene expression; and Pgp3, -5, and...

  1. Construction of a eukaryotic expression plasmid of Humanin*

    Luo, Ben-yan; Chen, Xiang-ming; Tang, Min; Chen, Feng; Chen, Zhi

    2004-01-01

    Objective: To construct a eukaryotic expression plasmid pcDNA3.1(-)-Humanin. Methods: The recombinant plasmid pGEMEX-1-Humanin was digested with restriction endonucleases BamH I and Hind III and the Humanin gene fragments, about 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1(-) and the recombinant plasmids pcDNA3.1(-)-Humanin were identified by sequencing. Results: Recombinant plasmid DNA successfully produced a band whic...

  2. Characterization of a plasmid from moderately halophilic eubacteria

    Fernández Castillo, Rosario; Vargas, C.; Nieto Gutiérrez, Joaquín José; Ventosa Ucero, Antonio; Ruiz Berraquero, Francisco

    1992-01-01

    A plasmid has been isolated for the first time from moderately halophilic eubacteria. Halomonas elongata, Halomonrrs halmphila, Deleya halophila and Vibrio costkola were found to harbour an 11.5 kbp plasmid (pMH1). The plasmid was isolated and characterized after transformation into Escherichia coh'JM101 cells. A restriction map was constructed, and unique restriction sites for EcoRI, EcoRV and ClaI were detected. The occurrence of such a plasmid in the original halophilic strains was confirm...

  3. Identification of plasmid partition function in coryneform bacteria

    Kurusu, Yasurou; Satoh, Yukie; Inui, Masayuki; Kohama, Keiko; Kobayashi, Miki; Terasawa, Masato; Yukawa, Hideaki (Mitsubishi Petrochemical Co., Ltd., Ibaraki (Japan))

    1991-03-01

    The authors have identified and characterized a partition function that is required for stable maintenance of plasmids in the coryneform bacteria Brevibacterium flavum MJ233 and Corynebacterium glutamicum ATCC 31831. This function is localized to a HindIII-NspV fragment (673 bp) adjacent to the replication region of the plasmid, named pBY503, from Brevibacterium stationis IFO 12144. The function was independent of copy number control and was not associated directly with plasmid replication functions. This fragment was able to stabilize the unstable plasmids in cis but not in trans.

  4. Plasmid P1 replication: negative control by repeated DNA sequences.

    Chattoraj, D; Cordes, K.; Abeles, A

    1984-01-01

    The incompatibility locus, incA, of the unit-copy plasmid P1 is contained within a fragment that is essentially a set of nine 19-base-pair repeats. One or more copies of the fragment destabilizes the plasmid when present in trans. Here we show that extra copies of incA interfere with plasmid DNA replication and that a deletion of most of incA increases plasmid copy number. Thus, incA is not essential for replication but is required for its control. When cloned in a high-copy-number vector, pi...

  5. Detection of an antioxidant profile in the human brain in vivo via double editing with MEGA-PRESS.

    Terpstra, Melissa; Marjanska, Malgorzata; Henry, Pierre-Gilles; Tkác, Ivan; Gruetter, Rolf

    2006-12-01

    Vitamin C (ascorbate) and glutathione (GSH) are the two most concentrated non-enzymatic antioxidants in the human brain. Double editing with (DEW) MEGA-PRESS at 4T was designed in this study to measure both antioxidants in the same amount of time previously required to measure one. In the occipital lobe of four human subjects, resolved ascorbate (Asc) and GSH resonances were detected repeatedly and simultaneously using DEW MEGA-PRESS. The Asc and GSH concentrations measured using LCModel analysis of DEW MEGA-PRESS spectra were 0.8 +/- 0.1 and 1.0 +/- 0.1 micromol/g (mean +/- SD), with average Cramer-Rao lower bounds (CRLB) of 10% and 7%, respectively. Aside from the effects of J-modulation at a common echo time (TE), double editing did not compromise sensitivity. To determine the extent to which the oxidized forms of Asc and GSH contribute to DEW MEGA-PRESS spectra in vivo, chemical shifts and coupling constants for dehydroascorbate (DHA) and oxidized glutathione (GSSG) were measured at physiologic pH and temperature. DHA does not contribute to the 3.73 ppm DEW MEGA-PRESS Asc resonance. GSSG contributions to the DEW MEGA-PRESS GSH resonance (3.0 ppm) are negligible under physiologic conditions, and would be evidenced by a distinct GSSG resonance (3.3 ppm) at exceptionally high concentrations. PMID:17089366

  6. Photonic plasmid stability of transformed Salmonella typhimurium: A comparison of three unique plasmids

    Acquiring a highly stable photonic plasmid in transformed Salmonella typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella typhimurium (S. typh-lux) u...

  7. Degradative Plasmid and Heavy Metal Resistance Plasmid Naturally Coexist in Phenol and Cyanide Assimilating Bacteria

    Bahig E.  Deeb; Abdullah D. Altalhi

    2009-01-01

    Problem statement: Heavy metals are known to be powerful inhibitors of xenobiotics biodegradation activities. Alleviation the inhibitory effect of these metals on the phenol biodegradation activities in presence of heavy metals resistant plasmid was investigated. Approach: Combination of genetic systems of degradation of xenobiotic compound and heavy metal resistance was one of the approaches to the creation of polyfunctional strains for bioremediation of s...

  8. Photonic plasmid stability of transformed Salmonella Typhimurium: A comparison of three unique plasmids

    Lay Donald

    2009-07-01

    Full Text Available Abstract Background Acquiring a highly stable photonic plasmid in transformed Salmonella Typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella Typhimurium (S. typh-lux using three different plasmids and characterize their respective photonic properties. Results In presence of ampicillin (AMP, S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 plasmids exhibited 100% photon-emitting colonies over a 10-d study period. Photon emitters of S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 without AMP selection decreased over time (P 7 to 1 × 109 CFU, P 0.05; although photonic emissions across a range of bacterial concentrations were not different (1 × 104 to 1 × 106 CFU, P > 0.05. For very low density bacterial concentrations imaged in 96 well plates photonic emissions were positively correlated with bacterial concentration (P 3 to 1 × 105 CFU low to high were different in the 96-well plate format (P Conclusion These data characterize photon stability properties for S. typh-lux transformed with three different photon generating plasmids that may facilitate real-time Salmonella tracking using in vivo or in situ biophotonic paradigms.

  9. Photonic Plasmid Stability of Transformed Salmonella Typhimurium: A Comparison of Three Unique Plasmids

    Background: Acquiring a highly stable photonic plasmid in transformed Salmonella Typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella Typhimurium (S....

  10. Bacterial Mitosis: ParM of Plasmid R1 Moves Plasmid DNA by an Actin-like Insertional Polymerization Mechanism

    Møller-Jensen, Jakob; Borch, Jonas; Dam, Mette;

    2003-01-01

    Bacterial DNA segregation takes place in an active and ordered fashion. In the case of Escherichia coli plasmid R1, the partitioning system (par) separates paired plasmid copies and moves them to opposite cell poles. Here we address the mechanism by which the three components of the R1 par system...... act together to generate the force required for plasmid movement during segregation. ParR protein binds cooperatively to the centromeric parC DNA region, thereby forming a complex that interacts with the filament-forming actin-like ParM protein in an ATP-dependent manner, suggesting that plasmid...... movement is powered by insertional polymerization of ParM. Consistently, we find that segregating plasmids are positioned at the ends of extending ParM filaments. Thus, the process of R1 plasmid segregation in E. coli appears to be mechanistically analogous to the actin-based motility operating...

  11. Mega clinical trials which have shaped the RAS intervention clinical practice.

    Düsing, Rainer

    2016-06-01

    Following extensive clinical research, drugs affecting the renin-angiotensin system have been used for the treatment of patients with congestive heart failure, myocardial infarction, hypertension, diabetic nephropathy, chronic renal failure and for reducing the risk of developing major cardiovascular (CV) events. This review examines all mega trials (those involving >1000 patients) and smaller pivotal trials involving angiotensin-converting enzyme inhibitors (ACE-Is; 25 mega trials) and angiotensin receptor blockers (ARBs; 27 mega trials) to provide perspective on the huge database of evidence that has accumulated on the use of these drugs. Our review demonstrates that ACE-Is and ARBs are generally as effective as conventional therapies in the treatment of hypertension, but offer additional cardioprotective benefits in patients with heart failure, and in those who have experienced myocardial infarction. Also, both ACE-Is and ARBs are capable of renal protection in addition to their blood-pressure-lowering effects. Although ACE-Is and ARBs provide major benefits to CV patients, doubts remain over the concept of blood-pressure-independent CV protection offered by both classes of drugs. ACE-Is and ARBs appear to be equally effective with respect to morbidity and mortality endpoints, but ARBs are better tolerated. Considering the available evidence, the combined use of an ACE-I and ARB should be avoided and full doses of either ACE-I or ARB should be aimed for as evidence suggests they provide a greater prognostic benefit. PMID:27271312

  12. MegaTevs: single-chain dual nucleases for efficient gene disruption.

    Wolfs, Jason M; DaSilva, Matthew; Meister, Sarah E; Wang, Xu; Schild-Poulter, Caroline; Edgell, David R

    2014-07-01

    Targeting gene disruptions in complex genomes relies on imprecise repair by the non-homologous end-joining DNA pathway, creating mutagenic insertions or deletions (indels) at the break point. DNA end-processing enzymes are often co-expressed with genome-editing nucleases to enhance the frequency of indels, as the compatible cohesive ends generated by the nucleases can be precisely repaired, leading to a cycle of cleavage and non-mutagenic repair. Here, we present an alternative strategy to bias repair toward gene disruption by fusing two different nuclease active sites from I-TevI (a GIY-YIG enzyme) and I-OnuI E2 (an engineered meganuclease) into a single polypeptide chain. In vitro, the MegaTev enzyme generates two double-strand breaks to excise an intervening 30-bp fragment. In HEK 293 cells, we observe a high frequency of gene disruption without co-expression of DNA end-processing enzymes. Deep sequencing of disrupted target sites revealed minimal processing, consistent with the MegaTev sequestering the double-strand breaks from the DNA repair machinery. Off-target profiling revealed no detectable cleavage at sites where the I-TevI CNNNG cleavage motif is not appropriately spaced from the I-OnuI binding site. The MegaTev enzyme represents a small, programmable nuclease platform for extremely specific genome-engineering applications. PMID:25013171

  13. Wireless Laser Range Finder System for Vertical Displacement Monitoring of Mega-Trusses during Construction

    Hyo Seon Park

    2013-05-01

    Full Text Available As buildings become increasingly complex, construction monitoring using various sensors is urgently needed for both more systematic and accurate safety management and high-quality productivity in construction. In this study, a monitoring system that is composed of a laser displacement sensor (LDS and a wireless sensor node was proposed and applied to an irregular building under construction. The subject building consists of large cross-sectional members, such as mega-columns, mega-trusses, and edge truss, which secured the large spaces. The mega-trusses and edge truss that support this large space are of the cantilever type. The vertical displacement occurring at the free end of these members was directly measured using an LDS. To validate the accuracy and reliability of the deflection data measured from the LDS, a total station was also employed as a sensor for comparison with the LDS. In addition, the numerical simulation result was compared with the deflection obtained from the LDS and total station. Based on these investigations, the proposed wireless displacement monitoring system was able to improve the construction quality by monitoring the real-time behavior of the structure, and the applicability of the proposed system to buildings under construction for the evaluation of structural safety was confirmed.

  14. Mega-heatwave temperatures due to combined soil desiccation and atmospheric heat accumulation

    Miralles, Diego G.; Teuling, Adriaan J.; van Heerwaarden, Chiel C.; Vilà-Guerau de Arellano, Jordi

    2014-05-01

    The recent European mega-heatwaves of 2003 and 2010 broke temperature records across Europe. Although events of this magnitude were unprecedented from a historical perspective, they are expected to become common by the end of the century. However, our understanding of extreme heatwave events is limited and their representation in climate models remains imperfect. Here we investigate the physical processes underlying recent mega-heatwaves using satellite and balloon measurements of land and atmospheric conditions from the summers of 2003 in France and 2010 in Russia, in combination with a soil-water-atmosphere model. We find that, in both events, persistent atmospheric pressure patterns induced land-atmosphere feedbacks that led to extreme temperatures. During daytime, heat was supplied by large-scale horizontal advection, warming of an increasingly desiccated land surface and enhanced entrainment of warm air into the atmospheric boundary layer. Overnight, the heat generated during the day was preserved in an anomalous kilometres-deep atmospheric layer located several hundred metres above the surface, available to re-enter the atmospheric boundary layer during the next diurnal cycle. This resulted in a progressive accumulation of heat over several days, which enhanced soil desiccation and led to further escalation in air temperatures. Our findings suggest that the extreme temperatures in mega-heatwaves can be explained by the combined multi-day memory of the land surface and the atmospheric boundary layer.

  15. The effects of the MEGA BORG spill on Gulf of Mexico shrimp

    A brown shrimp stock model was developed at the NMFS Galveston Laboratory to assess the effects of the MEGA BORG oil spill on the brown shrimp population along the Texas coast. Water and sediment samples from near the MEGA BORG spill area were collected and analyzed by the Geochemical and Environmental Research Group at Texas A ampersand M University. Hydrocarbon concentrations found at the sediment collection stations ranged from 1.38 ug/g to 7.42 ug/g, while concentrations at the water column sampling sites ranged from 0.69 ug/l to 27.39 ug/l. Evaluation of chromatograms and analytical data suggested that sediments from the area contained primarily biogenic hydrocarbons. Only one station had elevated hydrocarbon levels. However, these values were only three to five times higher than the concentrations measured at the other stations. Evaluation of the water samples from the area showed only very low concentration levels of hydrocarbons were present in the water column. A bioassay study was conducted by the US Environmental Protection Agency, Environmental Research Laboratory in Gulf Breeze, Florida. The main conclusion from the bioassay study was that the concentrations of hydrocarbons measured at the field sites where water samples were taken tended to be three orders of magnitude lower than the hydrocarbon concentrations causing toxic responses in mysid and white shrimp. There were no detected effects of the brown shrimp population along the Texas coast from the MEGA BORG oil spill

  16. Radar scattering of linear dunes and mega-yardangs: Application to Titan

    Paillou, Philippe; Radebaugh, Jani; Wall, Stephen

    2015-01-01

    The Ku-band (13.8 GHz - 2.2 cm) RADAR instrument onboard the Cassini-Huygens spacecraft has revealed the richness of the surface of Titan, as numerous seas, lakes, rivers, cryo-volcanic flows and vast dune fields have been discovered. Linear dunes are a major geomorphological feature present on Titan, covering up to 17% of its surface, mainly in equatorial regions. However, the resolution of the RADAR instrument is not good enough to allow a detailed study of the morphology of these features. In addition, other linear wind-related landforms, such as mega-yardangs (linear wind-abraded ridges formed in cohesive rocks), are likely to present a comparable radar signature that could be confused with the one of dunes. We conducted a comparative study of the radar radiometry of both linear dunes and mega-yardangs, based on representative terrestrial analogues: the linear dunes located in the Great Sand Sea in western Egypt and in the Namib Desert in Namibia, and the mega-yardangs observed in the Lut Desert in easter...

  17. Wireless laser range finder system for vertical displacement monitoring of mega-trusses during construction.

    Park, Hyo Seon; Son, Sewook; Choi, Se Woon; Kim, Yousok

    2013-01-01

    As buildings become increasingly complex, construction monitoring using various sensors is urgently needed for both more systematic and accurate safety management and high-quality productivity in construction. In this study, a monitoring system that is composed of a laser displacement sensor (LDS) and a wireless sensor node was proposed and applied to an irregular building under construction. The subject building consists of large cross-sectional members, such as mega-columns, mega-trusses, and edge truss, which secured the large spaces. The mega-trusses and edge truss that support this large space are of the cantilever type. The vertical displacement occurring at the free end of these members was directly measured using an LDS. To validate the accuracy and reliability of the deflection data measured from the LDS, a total station was also employed as a sensor for comparison with the LDS. In addition, the numerical simulation result was compared with the deflection obtained from the LDS and total station. Based on these investigations, the proposed wireless displacement monitoring system was able to improve the construction quality by monitoring the real-time behavior of the structure, and the applicability of the proposed system to buildings under construction for the evaluation of structural safety was confirmed. PMID:23648650

  18. Induced mutagenesis of plasmids and chromosomal genes inserted into plasmid DNA 1. Mutagenic effects of irradiations

    Effect of two physical agents: UV- and γ-radiation has been considered in comparison. DNA of RSF2124 plasmid, determining colcine synthesis and ampicillin resistance, was used as a model. Mutagenous effect is taken into account according to the appearance of Col--mutants, which are not capable of colicine synthesis. Lethal effect is determined according to ampicillin marker inactivation. After reisolation of plasmid DNA from mutant transformant, new traits and antibiotic resistance are preserved during subsequent transformations and reseedings of transformed colonies, which proves mutational nature of the transformations. Under short-wave UV irradiation (lambda=254 nm) of RSF2124 DNA a clear mutagenous effect is detected: relative amount of Col--mutants at the optimum for mutagenesis doses increased by a factor of 10. Under conditions of W-reactivation (additional UV-irradiation of recipient cells of wild C600 type) of lethal injuries an increase in mutagenous effect was observed, which is reliable for 95%. A distinct increase in mutagenesis (approximately by a factor of 4) is observed during UV-irradiation in small doses of only one recipient cell (a so-called indirect UV-mutagenesis). Thus, according to its ability to W- and indirect UV-mutagenesis plasmid DNA behaves as DNA of moderate phages, which can testify to their evolution relationship. Treatment of plasmid DNA with acridine orange before UV-irradiation protected only from lethal injuries. γ-irradiation of 60Co at inactivation approximately 10-2 increased by an order the yield of Col--mutants. The presence of the plasmid in a cell did not affect its UV-resistance

  19. Degradative Plasmid and Heavy Metal Resistance Plasmid Naturally Coexist in Phenol and Cyanide Assimilating Bacteria

    Bahig E.  Deeb

    2009-01-01

    Full Text Available Problem statement: Heavy metals are known to be powerful inhibitors of xenobiotics biodegradation activities. Alleviation the inhibitory effect of these metals on the phenol biodegradation activities in presence of heavy metals resistant plasmid was investigated. Approach: Combination of genetic systems of degradation of xenobiotic compound and heavy metal resistance was one of the approaches to the creation of polyfunctional strains for bioremediation of soil after co-contamination with organic pollutants and heavy metals. Results: A bacterial strain Pseudomonas putida PhCN (pPhCN1, pPhCN2 had been obtained. This bacterium contained two plasmids, a 120 Kb catabolic plasmid that encode for breakdown of phenol (pPhCN1 and pPhCN2 plasmid (100 Kb that code for cadmium and copper resistant. Cyanide assimilation by this bacterium was encoded by chromosomal genes. The inhibitory effect of cadmium (Cd2+ or copper (Cu2+ on the degradation of phenol and cyanide by P. putida strains PhCN and PhCN1 (contained pPhCN1 were investigated. The resistant strain PhCN showed high ability to degrade phenol and cyanide in presence of Cd2+ or Cu2+ comparing with the sensitive strain PhCN1. In addition, Cd2+ or Cu2+ was also found to exert a strong inhibitory effect on the C23O dioxygenase enzyme activity in the presence of cyanide as a nitrogen source. Conclusion: The presence of heavy metal resistance plasmid alleviated the inhibitory effect of metals on the phenol and cyanide assimilation by resistant strain.

  20. Mutations in an antisense RNA, involved in the replication control of a repABC plasmid, that disrupt plasmid incompatibility and mediate plasmid speciation.

    Rivera-Urbalejo, América; Pérez-Oseguera, Ángeles; Carreón-Rodríguez, Ofelia E; Cevallos, Miguel A

    2015-03-01

    The maintenance of large plasmid in a wide variety of alpha-proteobacteria depends on the repABC replication/segregation unit. The intergenic repB-repC region of these plasmids encodes a countertranscribed RNA (ctRNA) that modulates the transcription/translation rate of RepC, the initiator protein. The ctRNA acts as a strong incompatibility factor when expressed in trans. We followed a site directed mutagenesis approach to map those sequences of the ctRNA that are required for plasmid incompatibility and for plasmid replication control. We found that the first three nucleotides of the 5'-end of the ctRNA are essential for interactions with its target RNA. We also found that stretches of 4-5 nucleotides of non-complementarity within the first 10 nucleotides of the left arm of the ctRNA and the target RNA are sufficient to avoid plasmid incompatibility. Additionally, miniplasmid derivatives expressing ctRNAs with mutations in the 5' end or small deletions in the ctRNA are capable of controlling their own replication and coexisting with the parental plasmid. We suggest that a mechanism that could have a crucial role in the speciation process of repABC plasmids is to accumulate enough changes in this small region of the ctRNA gene to disrupt heteroduplex formation between the target RNA of one plasmid and the ctRNA of the other. Plasmids carrying these changes will not have defects in their maintenance. PMID:25644116

  1. Resident Exposure to Peripheral Nerve Surgical Procedures During Residency Training.

    Gil, Joseph A; Daniels, Alan H; Akelman, Edward

    2016-05-01

    Background Variability in case exposures has been identified for orthopaedic surgery residents. It is not known if this variability exists for peripheral nerve procedures. Objective The objective of this study was to assess ACGME case log data for graduating orthopaedic surgery, plastic surgery, general surgery, and neurological surgery residents for peripheral nerve surgical procedures and to evaluate intraspecialty and interspecialty variability in case volume. Methods Surgical case logs from 2009 to 2014 for the 4 specialties were compared for peripheral nerve surgery experience. Peripheral nerve case volume between specialties was performed utilizing a paired t test, 95% confidence intervals were calculated, and linear regression was calculated to assess the trends. Results The average number of peripheral nerve procedures performed per graduating resident was 54.2 for orthopaedic surgery residents, 62.8 for independent plastic surgery residents, 84.6 for integrated plastic surgery residents, 22.4 for neurological surgery residents, and 0.4 for surgery residents. Intraspecialty comparison of the 10th and 90th percentile peripheral nerve case volume in 2012 revealed remarkable variability in training. There was a 3.9-fold difference within orthopaedic surgery, a 5.0-fold difference within independent plastic surgery residents, an 8.8-fold difference for residents from integrated plastic surgery programs, and a 7.0-fold difference within the neurological surgery group. Conclusions There is interspecialty and intraspecialty variability in peripheral nerve surgery volume for orthopaedic, plastic, neurological, and general surgery residents. Caseload is not the sole determinant of training quality as mentorship, didactics, case breadth, and complexity play an important role in training. PMID:27168883

  2. Shared residence: lessons from Sweden

    Newnham, Annika

    2011-01-01

    This article, which draws on Swedish experience, argues that the English courts are currently using the shared residence order in ways that are unlikely either to benefit children or produce the intended results. Shared residence is considerably more demanding than co-parenting within an intact family, and attempts to use this order to improve parental co-operation are likely to prove counter productive. Using shared residence to send symbolic messages about parental status is not only contra...

  3. Construction and Application of R Prime Plasmids, Carrying Different Segments of an Octopine Ti Plasmid from Agrobacterium tumefaciens, for Complementation of vir Genes

    Hille, Jacques; Klasen, Ina; Schilperoort, Rob

    1982-01-01

    Several R prime plasmids have been obtained with high efficiency, by enclosing the R plasmid replicator, in an R::Ti cointegrate plasmid, between two copies of the transposon Tn1831, in the same orientation. These R primes carry different segments of an octopine Ti plasmid, and are compatible with T

  4. Homology of plasmids in strains of unicellular cyanobacteria

    Hondel, C.A.M.J.J. van den; Keegstra, W.; Borrias, W.E.; Arkel, G.A. van

    1979-01-01

    Six strains of unicellular cyanobacteria were examined for the presence of plasmids. Analysis of lysates of these strains by CsCl-ethidium bromide density centrifugation yielded a major chromosomal DNA band and a minor band containing covalently closed circular plasmid DNA, as shown by electron micr

  5. Transformation of Bacillus subtilis by single-stranded plasmid DNA.

    Rudolph, C F; Schmidt, B J; Saunders, C W

    1986-01-01

    The single-stranded form of a pE194-based plasmid transformed Bacillus subtilis protoplasts at least as efficiently as did the double-stranded plasmid, but the single-stranded form did not detectably transform B. subtilis competent cells.

  6. Curing of the 2 mu DNA plasmid from Saccharomyces cerevisiae.

    Toh-e, A; Wickner, R B

    1981-01-01

    The 2 mu DNA plasmid is often eliminated from yeast cells when they are transformed with the 2 mu DNA-LEU2-pMB9 composite plasmid pJDB219. Since pJDB219 is subsequently lost with high frequency, derivatives lacking all 2 mu DNA can be prepared from any strain.

  7. Purification of large plasmids with methacrylate monolithic columns.

    Krajnc, Nika Lendero; Smrekar, Franci; Cerne, Jasmina; Raspor, Peter; Modic, Martina; Krgovic, Danijela; Strancar, Ales; Podgornik, Ales

    2009-08-01

    The rapid evolution of gene therapy and DNA vaccines results in an increasing interest in producing large quantities of pharmaceutical grade plasmid DNA. Most current clinical trials involve plasmids of 10 kb or smaller in size, however, future requirements for multigene vectors including extensive control regions may require the production of larger plasmids, e. g., 20 kb and bigger. The objective of this study was to examine certain process conditions for purification of large plasmids with the size of up to 93 kb. Since there is a lack of knowledge about production and purification of bigger plasmid DNA, cell lysis and storage conditions were investigated. The impact of chromatographic system and methacrylate monolithic column on the degradation of plasmid molecules under nonbinding conditions at different flow rates was studied. Furthermore, capacity measurements varying salt concentration in loading buffer were performed and the capacities up to 13 mg of plasmid per mL of the monolithic column were obtained. The capacity flow independence in the range from 130 to 370 cm/h was observed. Using high resolution monolithic column the separation of linear and supercoiled isoforms of large plasmids was obtained. Last but not least, since the baseline separation of RNA and pDNA was achieved, the one step purification on larger CIM DEAE 8 mL tube monolithic column was performed and the fractions were analyzed by CIM analytical monolithic columns. PMID:19598166

  8. Production and pharmaceutical formulation of plasmid DNA vaccines

    van der Heijden, I.

    2013-01-01

    Research leading to the thesis ‘Production and pharmaceutical formulation of plasmid DNA vaccines‘ can be divided into two parts. The first part describes the development of a Good Manufacturing Practice (GMP) compliant plasmid DNA production process of pDNA vaccines for the treatment of Human papil

  9. Two novel conjugative plasmids from a single strain of Sulfolobus

    Erauso, G.; Stedman, K.M.; Werken, van de H.J.G.; Zillig, W.; Oost, van der J.

    2006-01-01

    Two conjugative plasmids (CPs) were isolated and characterized from the same 'Sulfolobus islandicus' strain, SOG2/4, The plasmids were separated from each other and transferred into Sulfolobus soltataricus. One has a high copy number and is not stable (pSOG1) whereas the other has a low copy number

  10. Inc A/C Plasmids in Multidrug resistant Salmonella enterica

    Bacteria plasmids are fragments of extra-chromosomal double stranded deoxyribonucleic acid (DNA) that can contain a variety of genes beneficial to the survival of the host bacteria. Classification and tracking of plasmids is beneficial because they are potentially a medium of horizontal gene transf...