79 resultados para random mutagenesis

em Queensland University of Technology - ePrints Archive


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An in vivo screen has been devised for NF-κB p50 activity in Escherichia coli exploiting the ability of the mammalian transcription factor to emulate a prokaryotic repressor. Active intracellular p50 was shown to repress the expression of a green fluorescent protein reporter gene allowing for visual screening of colonies expressing active p50 on agar plates. A library of mutants was constructed in which the residues Y267, L269, A308 and V310 of the dimer interface were simultaneously randomised and twenty-five novel functional interfaces were selected which repressed the reporter gene to similar levels as the wild-type protein. The leucine-269 alanine-308 core was repeatedly, but not exclusively, selected from the library whilst a diversity of predominantly non-polar residues were selected at positions 267 and 310. These results indicate that L269 and A308 may form a hot spot of interaction and allow an insight into the processes of dimer selectivity and evolution within this family of transcription factors.

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Channel measurements and simulations have been carried out to observe the effects of pedestrian movement on multiple-input multiple-output orthogonal frequency division multiplexing (MIMO-OFDM) channel capacity. An in-house built MIMO-OFDM packet transmission demonstrator equipped with four transmitters and four receivers has been utilized to perform channel measurements at 5.2 GHz. Variations in the channel capacity dynamic range have been analysed for 1 to 10 pedestrians and different antenna arrays (2 × 2, 3 × 3 and 4 × 4). Results show a predicted 5.5 bits/s/Hz and a measured 1.5 bits/s/Hz increment in the capacity dynamic range with the number of pedestrian and the number of antennas in the transmitter and receiver array.

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Random Indexing K-tree is the combination of two algorithms suited for large scale document clustering.

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In this paper, we consider a time-space fractional diffusion equation of distributed order (TSFDEDO). The TSFDEDO is obtained from the standard advection-dispersion equation by replacing the first-order time derivative by the Caputo fractional derivative of order α∈(0,1], the first-order and second-order space derivatives by the Riesz fractional derivatives of orders β 1∈(0,1) and β 2∈(1,2], respectively. We derive the fundamental solution for the TSFDEDO with an initial condition (TSFDEDO-IC). The fundamental solution can be interpreted as a spatial probability density function evolving in time. We also investigate a discrete random walk model based on an explicit finite difference approximation for the TSFDEDO-IC.

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This paper describes the approach taken to the clustering task at INEX 2009 by a group at the Queensland University of Technology. The Random Indexing (RI) K-tree has been used with a representation that is based on the semantic markup available in the INEX 2009 Wikipedia collection. The RI K-tree is a scalable approach to clustering large document collections. This approach has produced quality clustering when evaluated using two different methodologies.

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In cloud computing resource allocation and scheduling of multiple composite web services is an important challenge. This is especially so in a hybrid cloud where there may be some free resources available from private clouds but some fee-paying resources from public clouds. Meeting this challenge involves two classical computational problems. One is assigning resources to each of the tasks in the composite web service. The other is scheduling the allocated resources when each resource may be used by more than one task and may be needed at different points of time. In addition, we must consider Quality-of-Service issues, such as execution time and running costs. Existing approaches to resource allocation and scheduling in public clouds and grid computing are not applicable to this new problem. This paper presents a random-key genetic algorithm that solves new resource allocation and scheduling problem. Experimental results demonstrate the effectiveness and scalability of the algorithm.

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Non-pathogenic lactic acid bacteria are economically important Gram-positive bacteria used extensively in the food industry. Due to their “generally regarded as safe” status, certain species from the genera Lactobacillus and Lactococcus are also considered desirable as candidates for the production and secretion of recombinant proteins, particular those with therapeutic applications. The hypothesis examined by this thesis is that Lactococcus lactis can be modified to be an effective antimicrobial agent. Therefore, the aims of this thesis were to investigate the optimisation of the expression, secretion and/or activities of potential heterologous antimicrobial proteins by the model lactic acid bacterium, Lactococcus lactis subsp. cremoris MG1363. L. lactis strains were engineered to express and secrete the recombinant CyuC surface protein from Lactobacillus reuteri BR11, and a fusion protein consisting of CyuC and lysostaphin using the Sep promoter and secretion signal. CyuC has been characterised as a cystine-binding protein, but has also been demonstrated to have fibronectin binding activity. Lysostaphin is a bacteriolytic enzyme with specific activity against the Gram-positive pathogen, Staphylococcus aureus. These modified L. lactis strains were then investigated to see if they had the ability to inhibit the adhesion of S. aureus to host extracellular matrix (ECM) proteins. It was observed that the cell extracts of the L. lactis strain with the vector only (pGhost9:ISS1) was able to inhibit the adhesion of S. aureus to fibronectin, whilst the cell extracts of the L. lactis strain expressing lysostaphin was able to inhibit adhesion to keratin. Finally, this thesis has identified specific lactococcal genes that affect the secretion of lysostaphin through the use of random transposon mutagenesis. Ten mutants with higher lysostaphin activity contained insertions in four different genes encoding: (i) an uncharacterised putative transmembrane protein (llmg_0609), (ii) an enzyme catalysing the first step in peptidoglycan biosynthesis (murA2), (iii) a homolog of the oxidative defence regulator (trmA), and (iv) an uncharacterised putative enzyme involved in ubiquinone biosynthesis (llmg_2148). The higher lysostaphin activity observed in these mutants was found to be due to higher amounts of lysostaphin being secreted. The findings of this thesis contribute to the development of this organism as an antimicrobial agent and also to our understanding of L. lactis genetics.

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Quantitative studies of nascent entrepreneurs such as GEM and PSED are required to generate their samples by screening the adult population, usually by phone in developed economies. Phone survey research has recently been challenged by shifting patterns of ownership and response rates of landline versus mobile (cell) phones, particularly for younger respondents. This challenge is acutely intense for entrepreneurship which is a strongly age-dependent phenomenon. Although shifting ownership rates have received some attention, shifting response rates have remained largely unexplored. For the Australian GEM 2010 adult population study we conducted a dual-frame approach that allows comparison between samples of mobile and landline phones. We find a substantial response bias towards younger, male and metropolitan respondents for mobile phones – far greater than explained by ownership rates. We also found these response rate differences significantly biases the estimates of the prevalence of early stage entrepreneurship by both samples, even when each sample is weighted to match the Australian population.