870 resultados para antisense transformation


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Presently organisations engage in what is termed as Global Business Transformation Projects [GBTPs], for consolidating, innovating, transforming and restructuring their processes and business strategies while undergoing fundamental change. Culture plays an important role in global business transformation projects as these involve people of different cultural backgrounds and span across countries, industries and disciplinary boundaries. Nevertheless, there is scant empirical research on how culture is conceptualised beyond national and organisational cultures but also on how culture is to be taken into account and dealt with within global business transformation projects. This research is situated in a business context and discovers a theory that aids in describing and dealing with culture. It draws on the lived experiences of thirty-two senior management practitioners, reporting on more than sixty-one global business transformation projects in which they were actively involved. The research method used is a qualitative and interpretive one and applies a grounded theory approach, with rich data generated through interviews. In addition, vignettes were developed to illustrate the derived theoretical models. The findings from this study contribute to knowledge in multiple ways. First, it provides a holistic account of global business transformation projects that describe the construct of culture by the elements of culture types, cultural differences and cultural diversity. A typology of culture types has been developed which enlarges the view of culture beyond national and organisational culture including an industry culture, professional service firm culture and 'theme' culture. The amalgamation of the culture types instantiated in a global business transformation project compromises its project culture. Second, the empirically grounded process for managing culture in global business transformation projects integrates the stages of recognition, understanding and management as well as the enablement providing a roadmap for dealing with culture in global business transformation projects. Third, this study identified contextual variables to global business transformation projects, which provide the means of describing the environment global business transformation projects are situated, influence the construct of culture and inform the process for managing culture. Fourth, the contribution to the research method is the positioning of interview research as a strategy for data generation and the detailed documentation applying grounded theory to discover theory.

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Many aspects of China's academic publishing system differ from the systems found in liberal market based economies of the United States, Western Europe and Australia. A high level of government intervention in both the publishing industry and academia and the challenges associated with attempting to make a transition from a centrally controlled towards a more market based publishing industry are two notable differences; however, as in other countries, academic communities and publishers are being transformed by digital technologies. This research explores the complex yet dynamic digital transformation of academic publishing in China, with a specific focus of the open and networked initiatives inspired by Web 2.0 and social media. The thesis draws on two case studies: Science Paper Online, a government-operated online preprint platform and open access mandate; and New Science, a social reference management website operated by a group of young PhD students. Its analysis of the innovations, business models, operating strategies, influences, and difficulties faced by these two initiatives highlights important characteristics and trends in digital publishing experiments in China. The central argument of this thesis is that the open and collaborative possibilities of Web 2.0 inspired initiatives are emerging outside the established journal and monograph publishing system in China, introducing innovative and somewhat disruptive approaches to the certification, communication and commercial exploitation of knowledge. Moreover, emerging publishing models are enabling and encouraging a new system of practising and communicating science in China, putting into practice some elements of the Open Science ethos. There is evidence of both disruptive change to old publishing structures and the adaptive modification of emergent replacements in the Chinese practice. As such, the transformation from traditional to digital and interactive modes of publishing, involves both competition and convergence between new and old publishers, as well as dynamics of co-evolution involving new technologies, business models, social norms, and government reform agendas. One key concern driving this work is whether there are new opportunities and new models for academic publishing in the Web 2.0 age and social media environment, which might allow the basic functions of communication and certification to be achieved more effectively. This thesis enriches existing knowledge of open and networked transformations of scholarly publishing by adding a Chinese story. Although the development of open and networked publishing platforms in China remains in its infancy, the lessons provided by this research are relevant to practitioners and stakeholders interested in understanding the transformative dynamics of networked technologies for publishing and advocating open access in practice, not only in China, but also internationally.

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One set of public institutions that has seen growing discussion about the transformative impact of new media technologies has been universities. The higher education sector, historically one of the more venerable and stable areas of public life, is now the subject of almost continuous speculation about whether it can continue in its current form during the 21st century. Digital media technologies are often seen as being at the forefront of such changes. It has been widely noted that moves towards a knowledge economy generates ‘skills-biased technological change’, that places a premium upon higher education qualifications, and that this earnings gap remains despite the continuing increase in the number of university graduates. As the demand for higher education continues to grow worldwide, there are new discussions about whether technologically-mediated education through new forms such as Massively Open Online Courses (MOOCs) are broadening access to quality learning, or severing the vital connection between teacher and student seen as integral to the learning process. This paper critically appraises such debates in the context of early 21st century higher education. It will discuss ten drivers of change in higher education, many of which are related to themes discussed elsewhere in this book, such as the impact of social media, globalization, and a knowledge economy. It will also consider the issues raised in navigating such developments from the perspective of the ‘Five P’s’: practical issues; personal issues; pedagogical issues; policy issues; and philosophical issues. It also includes a critical evaluation of MOOCs from the point of view of their educational qualities. It will conclude with the observation that while universities will continue to play a significant – and perhaps growing – role in the economy, society and culture, the issues raised about what Clayton Christensen and Henry Eyring term the ‘disruptive university’ (Christensen and Eyring 2011) are nonetheless pressing ones, and that cost and policy pressures in particular are likely to generate significant institutional transformations in higher education worldwide.

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We demonstrate an unusual shape transformation of Ag nanospheres into {111}-oriented Au–Ag dendritic nanostructures by a galvanic replacement reaction in the ionic liquid (IL) 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIM][BF4]).

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The chemically reversible solid−solid phase transformation of a TCNQ-modified glassy carbon, indium tin oxide, or metal electrode into Co\[TCNQ]2(H2O)2 material in the presence of Co2+(aq) containing electrolytes has been induced and monitored electrochemically. Voltammetric data reveal that the TCNQ/Co\[TCNQ]2(H2O)2 interconversion process is independent of electrode material and identity of cobalt electrolyte anion. However, a marked dependence on electrolyte concentration, scan rate, and method of electrode modification (drop casting or mechanical attachment) is found. Cyclic voltammetric and double potential step chronoamperometric measurements confirm that formation of Co\[TCNQ]2(H2O)2 occurs through a rate-determining nucleation and growth process that initially involves incorporation of Co2+(aq) ions into the reduced TCNQ crystal lattice at the TCNQ|electrode|electrolyte interface. Similarly, the reverse (oxidation) process, which involves transformation of solid Co\[TCNQ]2(H2O)2 back to parent TCNQ crystals, also is controlled by nucleation−growth kinetics. The overall chemically reversible process that represents this transformation is described by the reaction:  2TCNQ0(s) + 2e- + Co2+(aq) + 2H2O \[Co(TCNQ)2(H2O)2](s). Ex situ SEM images illustrated that this reversible TCNQ/Co\[TCNQ]2(H2O)2 conversion process is accompanied by drastic size and morphology changes in the parent solid TCNQ. In addition, different sizes of needle-shaped nanorod/nanowire crystals of Co\[TCNQ]2(H2O)2 are formed depending on the method of surface immobilization.

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In situ atomic force microscopy (AFM) allows images from the upper face and sides of TCNQ crystals to be monitored during the course of the electrochemical solid–solid state conversion of 50 × 50 μm2 three-dimensional drop cast crystals of TCNQ to CuTCNQ or M[TCNQ]2(H2O)2 (M = Co, Ni). Ex situ images obtained by scanning electron microscopy (SEM) also allow the bottom face of the TCNQ crystals, in contact with the indium tin oxide or gold electrode surface and aqueous metal electrolyte solution, to be examined. Results show that by carefully controlling the reaction conditions, nearly mono-dispersed, rod-like phase I CuTCNQ or M[TCNQ]2(H2O)2 can be achieved on all faces. However, CuTCNQ has two different phases, and the transformation of rod-like phase 1 to rhombic-like phase 2 achieved under conditions of cyclic voltammetry was monitored in situ by AFM. The similarity of in situ AFM results with ex situ SEM studies accomplished previously implies that the morphology of the samples remains unchanged when the solvent environment is removed. In the process of crystal transformation, the triple phase solid∣electrode∣electrolyte junction is confirmed to be the initial nucleation site. Raman spectra and AFM images suggest that 100% interconversion is not always achieved, even after extended electrolysis of large 50 × 50 μm2 TCNQ crystals.

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The continuous growth of the XML data poses a great concern in the area of XML data management. The need for processing large amounts of XML data brings complications to many applications, such as information retrieval, data integration and many others. One way of simplifying this problem is to break the massive amount of data into smaller groups by application of clustering techniques. However, XML clustering is an intricate task that may involve the processing of both the structure and the content of XML data in order to identify similar XML data. This research presents four clustering methods, two methods utilizing the structure of XML documents and the other two utilizing both the structure and the content. The two structural clustering methods have different data models. One is based on a path model and other is based on a tree model. These methods employ rigid similarity measures which aim to identifying corresponding elements between documents with different or similar underlying structure. The two clustering methods that utilize both the structural and content information vary in terms of how the structure and content similarity are combined. One clustering method calculates the document similarity by using a linear weighting combination strategy of structure and content similarities. The content similarity in this clustering method is based on a semantic kernel. The other method calculates the distance between documents by a non-linear combination of the structure and content of XML documents using a semantic kernel. Empirical analysis shows that the structure-only clustering method based on the tree model is more scalable than the structure-only clustering method based on the path model as the tree similarity measure for the tree model does not need to visit the parents of an element many times. Experimental results also show that the clustering methods perform better with the inclusion of the content information on most test document collections. To further the research, the structural clustering method based on tree model is extended and employed in XML transformation. The results from the experiments show that the proposed transformation process is faster than the traditional transformation system that translates and converts the source XML documents sequentially. Also, the schema matching process of XML transformation produces a better matching result in a shorter time.

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While over the past decade many Australian schools have come to understand the transformative potential of digitally-rich teaching and learning, traditional models of schooling continue to dominate. Even with significant investment in the area, both in terms of digital resourcing and teacher professional development, innovation has generally only occurred in individual classrooms or ‘pockets’ in schools. This article discusses three interdependent conditions which need to exist as a foundation in order to facilitate the opportunity for transformation from traditional to digitally-rich ways of working in primary, middle and secondary schools or colleges. Distributed and transformational leadership approaches are critiqued with core elements identified which facilitate change. The establishment of a vision is identified and discussed as a fundamental driver and rudder for school transformation. The importance of creating and maintaining urgency to compel a school community to adopt and embed change is unpacked. This report concludes with a synthesis of the three preconditions and recommendations for proponents of digital school transformation.

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Many examples of extreme virus resistance and posttranscriptional gene silencing of endogenous or reporter genes have been described in transgenic plants containing sense or antisense transgenes. In these cases of either cosuppression or antisense suppression, there appears to be induction of a surveillance system within the plant that specifically degrades both the transgene and target RNAs. We show that transforming plants with virus or reporter gene constructs that produce RNAs capable of duplex formation confer virus immunity or gene silencing on the plants. This was accomplished by using transcripts from one sense gene and one antisense gene colocated in the plant genome, a single transcript that has self-complementarity, or sense and antisense transcripts from genes brought together by crossing. A model is presented that is consistent with our data and those of other workers, describing the processes of induction and execution of posttranscriptional gene silencing.

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Rice ragged stunt virus (RRSV) is an important pathogen of rice affecting its cultivation in South and South East Asia. An approach based on pathogen derived resistance (PDR) was used to produce RRSV resistant rice cultivars. Sequences from the coding region of RRSV genome segments 7 and 10 (non-structural genes), and 5, 8 and 9 (structural genes) were placed in sense or antisense orientation behind the plant expression promoters CaMV35S, RolC, Ubil, Actl and RBTV. Rice cultivars Taipei 309 and Chinsurah Boro II were transformed by biolistic and/or Agrobacterium-mediated delivery of one or more of these PDR gene constructs. A large number of transgenic lines were produced from calli derived from mature or immature embryos, co-bombarded with the marker gene hph encoding hygromycin resistance and RRSV PDR genes or co-cultivated with strains having the binary vector containing these two genes. Both Mendelian and non-Mendelian segregations were observed in transgenic progeny, especially with transgenic lines produced by biolistics. Preliminary tests conducted in China on selected transgenic lines indicate that plants with RRSV segment 5 antisense PDR gene confer RRSV resistance.

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A series of improved vectors have been constructed that are suitable for use in Agrobacterium tumefaciens-mediated monocot transformation. These binary vectors have several useful features, including the selectable marker genes bar (phosphinothricin resistance) or hph (hygromycin resistance) driven by either the cauliflower mosaic virus (CaMV) 35S promoter or the maize ubiquitin promoter, a high-copy-number replication origin that allows reliable mini-prep DNA isolation from Escherichia coli, and a polylinker sequence into which target genes can be easily inserted. A significant improvement has been made to the hph gene by the introduction of an intron into its coding region. The presence of the intron abolishes hph expression in A. tumefaciens, rendering the bacterium susceptible to the selective agent hygromycin B. The use of such an intron-hph vector thus enables the antibiotic in plant culture media to function as both a selective agent for transformed tissue and as a contraselective agent for A. tumefaciens growth, thus minimising the overgrowth of A. tumefaciens on plant tissues during transformation. Furthermore, the intron appears to be correctly spliced in plant cells and significantly enhances hph expression in transformed rice tissue. In our experiments, the use of the intron-hph vector increased the frequency of rice transformation and has enabled the production of transgenic barley.

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Efficient transformation of barley cv. Schooner was achieved using Agrobacterium delivery, hygromycin or bialaphos selection and embryogenic callus. Using this system, transgenic plants were generated that contained either the green fluorescent protein gene, or transgenes derived from barley yellow dwarf (BYDV) and cereal yellow dwarf (CYDV) viruses. Many of these plants contained 1-3 transgene copies that were inherited in a simple Mendelian manner. Some plants containing BYDV and/or CYDV derived transgenes showed reduced virus symptoms and rates of viral replication when challenged with the appropriate virus. The ability to transform Schooner is a significant advance for the Australian barley industry, as this elite malting variety is, and has for the last 15 years been, the most widely grown barley variety in eastern Australia.

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Post-transcriptional silencing of plant genes using anti-sense or co-suppression constructs usually results in only a modest proportion of silenced individuals. Recent work has demonstrated the potential for constructs encoding self-complementary 'hairpin' RNA (hpRNA) to efficiently silence genes. In this study we examine design rules for efficient gene silencing, in terms of both the proportion of independent transgenic plants showing silencing, and the degree of silencing. Using hpRNA constructs containing sense/anti-sense arms ranging from 98 to 853 nt gave efficient silencing in a wide range of plant species, and inclusion of an intron in these constructs had a consistently enhancing effect. Intron-containing constructs (ihpRNA) generally gave 90-100% of independent transgenic plants showing silencing. The degree of silencing with these constructs was much greater than that obtained using either co-suppression or anti-sense constructs. We have made a generic vector, pHANNIBAL, that allows a simple, single PCR product from a gene of interest to be easily converted into a highly effective ihpRNA silencing construct. We have also created a high-throughput vector, pHELLSGATE, that should facilitate the cloning of gene libraries or large numbers of defined genes, such as those in EST collections, using an in vitro recombinase system. This system may facilitate the large-scale determination and discovery of plant gene functions in the same way as RNAi is being used to examine gene function in Caenorhabditis elegans.

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We have tested a methodology for the elimination of the selectable marker gene after Agrobacterium-mediated transformation of barley. This involves segregation of the selectable marker gene away from the gene of interest following co-transformation using a plasmid carrying two T-DNAs, which were located adjacent to each other with no intervening region. A standard binary transformation vector was modified by insertion of a small section composed of an additional left and right T-DNA border, so that the selectable marker gene and the site for insertion of the gene of interest (GOI) were each flanked by a left and right border. Using this vector three different GOIs were transformed into barley. Analysis of transgene inheritance was facilitated by a novel and rapid assay utilizing PCR amplification from macerated leaf tissue. Co-insertion was observed in two thirds of transformants, and among these approximately one quarter had transgene inserts which segregated in the next generation to yield selectable marker-free transgenic plants. Insertion of non-T-DNA plasmid sequences was observed in only one of fourteen SMF lines tested. This technique thus provides a workable system for generating transgenic barley free from selectable marker genes, thereby obviating public concerns regarding proliferation of these genes.

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Two transgenic callus lines of rice, stably expressing a β-glucuronidase (GUS) gene, were supertransformed with a set of constructs designed to silence the resident GUS gene. An inverted-repeat (i/r) GUS construct, designed to produce mRNA with self-complementarity, was much more effective than simple sense and antisense constructs at inducing silencing. Supertransforming rice calluses with a direct-repeat (d/r) construct, although not as effective as those with the i/r construct, was also substantially more effective in silencing the resident GUS gene than the simple sense and antisense constructs. DNA hybridisation analyses revealed that every callus line supertransformed with either simple sense or antisense constructs, and subsequently showing GUS silencing, had the silence-inducing transgenes integrated into the plant genome in inverted-repeat configurations. The silenced lines containing i/r and d/r constructs did not necessarily have inverted-repeat T-DNA insertions. There was significant methylation of the GUS sequences in most of the silenced lines but not in the unsilenced lines. However, demethylation treatment of silenced lines with 5-azacytidine did not reverse the post-transcriptional gene silencing (PTGS) of GUS. Whereas the levels of RNA specific to the resident GUS gene were uniformly low in the silenced lines, RNA specific to the inducer transgenes accumulated to a substantial level, and the majority of the i/r RNA was unpolyadenylated. Altogether, these results suggest that both sense- and antisense-mediated gene suppression share a similar molecular basis, that unpolyadenylated RNA plays an important role in PTGS, and that methylation is not essential for PTGS.