947 resultados para site-directed mutagenesis
Resumo:
Much has been written on Off-site Manufacture (OSM) in construction, particularly regarding the perceived benefits and barriers to implementation. However, very little understanding of the state of OSM in the Australian construction industry exists. A ‘scoping study' has recently been undertaken to determine the ‘state-of-the-art’ of OSM in Australia. This involved several industry workshops, interviews and case studies across four major states of Australia. The study surveyed a range of suppliers across the construction supply-chain, incorporating the civil, commercial and housing segments of the market. This revealed that skills shortages and lack of adequate OSM knowledge are generally the greatest issues facing OSM in Australia. The drivers and constraints that emerged from the research were, in large measure, consistent with those found in the US and UK, although some Australian anomalies are evident, such as the geographical disparity of markets. A comparative analysis with similar studies in the UK and US is reported, illustrating both the drivers and constraints confronting the industry in Australia. OSM uptake into the future is however dependent on many factors, not least of which is a better understanding of the construction process and its associated costs.
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In 2004, with the increasing overloading restriction requirements of society in Anhui, a provincial comprehensive overloading transportation survey has been developed to take evaluations on overloading actuality and enforcement efficiency with the support of the World Bank. A total of six site surveys were conducted at Hefei, Fuyang, Luan, Wuhu, Huainan and Huangshan Areas with four main contents respectively: traffic volume, axle load, freight information and registration information. Via statistical analysis on the survey data, conclusions were gained that: vehicle overloading are very universal and serious problems at arterial highways in Anhui now. The traffic loads have far exceeded the designed endure capacity of highways and have caused prevalent premature pavement damage, especially for rigid pavement. The overloading trucks are unimpeded engaged in highway freight transportation actually due to the disordered overloading enforcement strategies and the deficient inspecting technologies.
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The Internet theoretically enables marketers to personalize a Website to an individual consumer. This article examines optimal Website design from the perspective of personality trait theory and resource-matching theory. The influence of two traits relevant to Internet Web-site processing—sensation seeking and need for cognition— were studied in the context of resource matching and different levels of Web-site complexity. Data were collected at two points of time: personality-trait data and a laboratory experiment using constructed Web sites. Results reveal that (a) subjects prefer Web sites of a medium level of complexity, rather than high or low complexity; (b)high sensation seekers prefer complex visual designs, and low sensation seekers simple visual designs, both in Web sites of medium complexity; and (c) high need-for-cognition subjects evaluated Web sites with high verbal and low visual complexity more favourably.
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The interaction of quercetin, which is a bioflavonoid, with bovine serum albumin (BSA) was investigated under pseudo-physiological conditions by the application of UV–vis spectrometry, spectrofluorimetry and cyclic voltammetry (CV). These studies indicated a cooperative interaction between the quercetin–BSA complex and warfarin, which produced a ternary complex, quercetin–BSA–warfarin. It was found that both quercetin and warfarin were located in site I. However, the spectra of these three components overlapped and the chemometrics method – multivariate curve resolution-alternating least squares (MCR-ALS) was applied to resolve the spectra. The resolved spectra of quercetin–BSA and warfarin agreed well with their measured spectra, and importantly, the spectrum of the quercetin–BSA–warfarin complex was extracted. These results allowed the rationalization of the behaviour of the overlapping spectra. At lower concentrations ([warfarin] < 1 × 10−5 mol L−1), most of the site marker reacted with the quercetin–BSA, but free warfarin was present at higher concentrations. Interestingly, the ratio between quercetin–BSA and warfarin was found to be 1:2, suggesting a quercetin–BSA–(warfarin)2 complex, and the estimated equilibrium constant was 1.4 × 1011 M−2. The results suggest that at low concentrations, warfarin binds at the high-affinity sites (HAS), while low-affinity binding sites (LAS) are occupied at higher concentrations.
Resumo:
Plants have been identified as promising expression systems for the commercial production of recombinant proteins. Plant-based protein production or “biofarming” offers a number of advantages over traditional expression systems in terms of scale of production, the capacity for post-translation processing, providing a product free of contaminants and cost effectiveness. A number of pharmaceutically important and commercially valuable proteins, such as antibodies, biopharmaceuticals and industrial enzymes are currently being produced in plant expression systems. However, several challenges still remain to improve recombinant protein yield with no ill effect on the host plant. The ability for transgenic plants to produce foreign proteins at commercially viable levels can be directly related to the level and cell specificity of the selected promoter driving the transgene. The accumulation of recombinant proteins may be controlled by a tissue-specific, developmentally-regulated or chemically-inducible promoter such that expression of recombinant proteins can be spatially- or temporally- controlled. The strict control of gene expression is particularly useful for proteins that are considered toxic and whose expression is likely to have a detrimental effect on plant growth. To date, the most commonly used promoter in plant biotechnology is the cauliflower mosaic virus (CaMV) 35S promoter which is used to drive strong, constitutive transgene expression in most organs of transgenic plants. Of particular interest to researchers in the Centre for Tropical Crops and Biocommodities at QUT are tissue-specific promoters for the accumulation of foreign proteins in the roots, seeds and fruit of various plant species, including tobacco, banana and sugarcane. Therefore this Masters project aimed to isolate and characterise root- and seed-specific promoters for the control of genes encoding recombinant proteins in plant-based expression systems. Additionally, the effects of matching cognate terminators with their respective gene promoters were assessed. The Arabidopsis root promoters ARSK1 and EIR1 were selected from the literature based on their reported limited root expression profiles. Both promoters were analysed using the PlantCARE database to identify putative motifs or cis-acting elements that may be associated with this activity. A number of motifs were identified in the ARSK1 promoter region including, WUN (wound-inducible), MBS (MYB binding site), Skn-1, and a RY core element (seed-specific) and in the EIR1 promoter region including, Skn-1 (seed-specific), Box-W1 (fungal elicitor), Aux-RR core (auxin response) and ABRE (ABA response). However, no previously reported root-specific cis-acting elements were observed in either promoter region. To confirm root specificity, both promoters, and truncated versions, were fused to the GUS reporter gene and the expression cassette introduced into Arabidopsis via Agrobacterium-mediated transformation. Despite the reported tissue-specific nature of these promoters, both upstream regulatory regions directed constitutive GUS expression in all transgenic plants. Further, similar levels of GUS expression from the ARSK1 promoter were directed by the control CaMV 35S promoter. The truncated version of the EIR1 promoter (1.2 Kb) showed some differences in the level of GUS expression compared to the 2.2 Kb promoter. Therefore, this suggests an enhancer element is contained in the 2.2 Kb upstream region that increases transgene expression. The Arabidopsis seed-specific genes ATS1 and ATS3 were selected from the literature based on their seed-specific expression profiles and gene expression confirmed in this study as seed-specific by RT-PCR analysis. The selected promoter regions were analysed using the PlantCARE database in order to identify any putative cis elements. The seed-specific motifs GCN4 and Skn-1 were identified in both promoter regions that are associated with elevated expression levels in the endosperm. Additionaly, the seed-specific RY element and the ABRE were located in the ATS1 promoter. Both promoters were fused to the GUS reporter gene and used to transform Arabidopsis plants. GUS expression from the putative promoters was consitutive in all transgenic Arabidopsis tissue tested. Importantly, the positive control FAE1 seed-specific promoter also directed constitutive GUS expression throughout transgenic Arabidopsis plants. The constitutive nature seen in all of the promoters used in this study was not anticipated. While variations in promoter activity can be caused by a number of influencing factors, the variation in promoter activity observed here would imply a major contributing factor common to all plant expression cassettes tested. All promoter constructs generated in this study were based on the binary vector pCAMBIA2300. This vector contains the plant selection gene (NPTII) under the transcriptional control of the duplicated CaMV 35S promoter. This CaMV 35S promoter contains two enhancer domains that confer strong, constitutive expression of the selection gene and is located immediately upstream of the promoter-GUS fusion. During the course of this project, Yoo et al. (2005) reported that transgene expression is significantly affected when the expression cassette is located on the same T-DNA as the 35S enhancer. It was concluded, the trans-acting effects of the enhancer activate and control transgene expression causing irregular expression patterns. This phenomenon seems the most plausible reason for the constitutive expression profiles observed with the root- and seed-specific promoters assessed in this study. The expression from some promoters can be influenced by their cognate terminator sequences. Therefore, the Arabidopsis ARSK1, EIR1, ATS1 and ATS3 terminator sequences were isolated and incorporated into expression cassettes containing the GUS reporter gene under the control of their cognate promoters. Again, unrestricted GUS activity was displayed throughout transgenic plants transformed with these reporter gene fusions. As previously discussed constitutive GUS expression was most likely due to the trans-acting effect of the upstream CaMV 35S promoter in the selection cassette located on the same T-DNA. The results obtained in this study make it impossible to assess the influence matching terminators with their cognate promoters have on transgene expression profiles. The obvious future direction of research continuing from this study would be to transform pBIN-based promoter-GUS fusions (ie. constructs containing no CaMV 35S promoter driving the plant selection gene) into Arabidopsis in order to determine the true tissue specificity of these promoters and evaluate the effects of their cognate 3’ terminator sequences. Further, promoter truncations based around the cis-elements identified here may assist in determining whether these motifs are in fact involved in the overall activity of the promoter.
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The creative practice: the adaptation of picture book The Empty City (Megarrity/Oxlade, Hachette 2007) into an innovative, interdisciplinary performance for children which combines live performance, music, projected animation and performing objects. The researcher, in the combined roles of writer/composer proposes deliberate experiments in music, narrative and emotion in the various drafts of the adaptation, and tests them in process and performance product. A particular method of composing music for live performance is tested in against the emergent needs of a collaborative, intermedial process. The unpredictable site of research means that this project is both looking to address both pre-determined and emerging points of inquiry. This analysis (directed by audience reception) finds that critical incidents of intermediality between music, narrative, action and emotion translate directly into highlights of the performance.
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RatSLAM is a system for vision based Simultaneous Localization and Mapping (SLAM) that has been shown to be capable of building stable representations of real world environments. In this paper we describe a method for using RatSLAM representations as the basis for navigation to designated goal locations. The method uses a new component, goal memory, to learn the temporal gradient between places. Paths are recalled or inferred from the goal memory by following the temporal gradient from the robot’s current position to the goal location. Experimental results have been gathered in a combined office and laboratory environment using a Pioneer robot. The experiments show that the robot can perform vision based SLAM on-line and in real time, and then use those representations immediately to navigate directly to designated goal locations.
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In 2009, Religious Education is a designated key learning area in Catholic schools in the Archdiocese of Brisbane and, indeed, across Australia. Over the years, though, different conceptualisations of the nature and purpose of religious education have led to the construction of different approaches to the classroom teaching of religion. By investigating the development of religious education policy in the Archdiocese of Brisbane from 1984 to 2003, the study seeks to trace the emergence of new discourses on religious education. The study understands religious education to refer to a lifelong process that occurs through a variety of forms (Moran, 1989). In Catholic schools, it refers both to co-curricula activities, such as retreats and school liturgies, and the classroom teaching of religion. It is the policy framework for the classroom teaching of religion that this study explores. The research was undertaken using a policy case study approach to gain a detailed understanding of how new conceptualisations of religious education emerged at a particular site of policy production, in this case, the Archdiocese of Brisbane. The study draws upon Yeatman’s (1998) description of policy as occurring “when social actors think about what they are doing and why in relation to different and alternative possible futures” (p. 19) and views policy as consisting of more than texts themselves. Policy texts result from struggles over meaning (Taylor, 2004) in which specific discourses are mobilised to support particular views. The study has a particular interest in the analysis of Brisbane religious education policy texts, the discursive practices that surrounded them, and the contexts in which they arose. Policy texts are conceptualised in the study as representing “temporary settlements” (Gale, 1999). Such settlements are asymmetrical, temporary and dependent on context: asymmetrical in that dominant actors are favoured; temporary because dominant actors are always under challenge by other actors in the policy arena; and context - dependent because new situations require new settlements. To investigate the official policy documents, the study used Critical Discourse Analysis (hereafter referred to as CDA) as a research tool that affords the opportunity for researchers to map and chart the emergence of new discourses within the policy arena. As developed by Fairclough (2001), CDA is a three-dimensional application of critical analysis to language. In the Brisbane religious education arena, policy texts formed a genre chain (Fairclough, 2004; Taylor, 2004) which was a focus of the study. There are two features of texts that form genre chains: texts are systematically linked to one another; and, systematic relations of recontextualisation exist between the texts. Fairclough’s (2005) concepts of “imaginary space” and “frameworks for action” (p. 65) within the policy arena were applied to the Brisbane policy arena to investigate the relationship between policy statements and subsequent guidelines documents. Five key findings emerged from the study. First, application of CDA to policy documents revealed that a fundamental reconceptualisation of the nature and purpose of classroom religious education in Catholic schools occurred in the Brisbane policy arena over the last twenty-five years. Second, a disjuncture existed between catechetical discourses that continued to shape religious education policy statements, and educational discourses that increasingly shaped guidelines documents. Third, recontextualisation between policy documents was evident and dependent on the particular context in which religious education occurred. Fourth, at subsequent links in the chain, actors created their own “imaginary space”, thereby altering orders of discourse within the policy arena, with different actors being either foregrounded or marginalised. Fifth, intertextuality was more evident in the later links in the genre chain (i.e. 1994 policy statement and 1997 guidelines document) than in earlier documents. On the basis of the findings of the study, six recommendations are made. First, the institutional Church should carefully consider the contribution that the Catholic school can make to the overall pastoral mission of the diocese in twenty-first century Australia. Second, policymakers should articulate a nuanced understanding of the relationship between catechesis and education with regard to the religion classroom. Third, there should be greater awareness of the connections among policies relating to Catholic schools – especially the connection between enrolment policy and religious education policy. Fourth, there should be greater consistency between policy documents. Fifth, policy documents should be helpful for those to whom they are directed (i.e. Catholic schools, teachers). Sixth, “imaginary space” (Fairclough, 2005) in policy documents needs to be constructed in a way that allows for multiple “frameworks for action” (Fairclough, 2005) through recontextualisation. The findings of this study are significant in a number of ways. For religious educators, the study highlights the need to develop a shared understanding of the nature and purpose of classroom religious education. It argues that this understanding must take into account the multifaith nature of Australian society and the changing social composition of Catholic schools themselves. Greater recognition should be given to the contribution that religious studies courses such as Study of Religion make to the overall religious development of a person. In view of the social composition of Catholic schools, there is also an issue of ecclesiological significance concerning the conceptualisation of the relationship between the institutional Catholic Church and Catholic schools. Finally, the study is of significance because of its application of CDA to religious education policy documents. Use of CDA reveals the foregrounding, marginalising, or excluding of various actors in the policy arena.
Resumo:
In the past few years, numerous data collection protocols have been developed for wireless sensor networks (WSNs). However, there has been no comparison of their relative performance in realistic environments. Here we report the results of an empirical study using a Fleck3 sensor network testbed for four different data collection protocols: One phase pull Directed Diffusion (DD), Expected Number of Transmissions (ETX), ETX with explicit acknowledgment (ETX-eAck), and ETX with implicit acknowledgment (ETX-iAck). Our empirical study provides useful insights for future sensor network deployments. When the required application end-to-end reliability is not strict (e.g., 70%) and link quality is good, DD and ETX are the best options because of their simplicity and low routing overhead. Both ETX-eAck and ETX-iAck achieve more than 90% end-to-end reliability when the link quality is reasonable (less than 25% packet loss). When the link quality is good, ETX-iAck introduces significantly less routing overhead (up to 50%) than ETX-eAck. However, if the radio transceiver supports variable packet length, ETX-eAck can outperform ETX-iAck when the link quality is poor. The important message from this paper is that choice of data collection protocol should come after the operating environment is understood. This understanding must include the characteristics of the radio transceiver, and link loss statistics from a long-term (across seasons and weather variation) radio survey of the site.
Resumo:
The controversial love affair of CS Lewis, Oxford scholar and writer of the Narnia Chronicles, is set in a constellation of music, sculpture and mime. CS Lewis’s intriguing relationship with poet Joy Davidman moves, inspires and confronts us with the big questions. Beauty contrasts with the ephemeral land of the shadows. Crossbow’s adaption of William Nicholson’s soulful and witty play explores the joy and the grief of “experience: that most brutal of teachers.” Showcasing the abilities of Brisbane and Sydney actors, the company that brought you The Miracle Worker and Anne of the Thousand Days, will quicken your senses and stir your heart with Shadowlands. All performances have a tactile tour of the stage 20 minutes before the start time of the show. Special signed performance for hearing impaired patrons Thur 5 Aug 2pm