92 resultados para Overexpression cassette construction


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We constructed a genomic DNA library for Lipotes vexillifer (L. vexillifer), the Baiji or Yangtze River dolphin, one of the most endangered mammals in the world. The library consists of 149,000 BAC clones, with an average insert size of 83 kb, representing approximately 3.4 haploid genome equivalents. PCR amplification of four known L. vexillifer genes yielded two to four positive clones each. To demonstrate the utility of this library, we isolated and sequenced the L. vexillifer alpha lactalbumin gene, which is a gene specific to mammals and one which has been widely used as molecular tool in phylogenetic analysis. We also end-sequenced 20 randomly selected clones, resulting in the identification of at least five new L. vexilliter genes, five SSR loci, and one SINE locus. These results suggest that this library is a valuable resource for candidate gene cloning, physical mapping, and genome sequencing of this important and threatened species.

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An antibody phage display library against White Spot Syndrome Virus (WSSV) was constructed. After four rounds of panning against WSSV, 192 out of 480 clones displayed WSSV binding activity. One of the positive clones, designated A1, had relatively higher activity specifically binding to WSSV A1-soluble, single-chain fragment variable (scFv) antibody has an affinity constant (K-aff) of 2.02 +/- 0.42 x 10(9) M-1. Dot blot assays showed that A1-soluble scFv could detect WSSV directly from shrimp hemolymph after 24-h feeding infection by WSSV. A1 scFv has potential for the development of a cheap, simple and sensitive diagnostic kit for WSSV in the field. (C) 2003 Elsevier Science B.V. All rights reserved.

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A mutant of Anabaena sp. strain PCC7120 requiring high CO2 was generated using Tn5 mutagenesis. This is the first data for a filamentous cyanobacterium. The mutant was capable of growing at 5% CO2, but incapable of growing at air levels of CO2. Southern hybridization analysis indicated that the Anabaena genome was inserted by the transposon at one site. The apparent photosynthetic affinity of the mutant to external dissolved inorganic carbon (DIC) was about 300 times lower that of the wild type (WT), and the medium alkalization rate as well as the carboxysomal carbonic anhydrase activity of the mutant was also lower than those of the WT. When the mutant was transferred from the culture medium bubbled with 5% CO2 to higher DIC (8.4% CO2) or 1% CO2, it showed similar responses to the WT. However, aberrant carboxysomes were found in the mutant cells through ultrastructural analysis, indicating it was most probably the wrong organization of the carboxysomes that eventually led to the inefficient operation of carboxysomal carbonic anhydrase and the subsequent defectiveness of the mutant in utilizing DIC.

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Since its completion in 1973 the Danjiangkou Dam has markedly changed downstream flows, water levels, temperatures, sediment loads and other water quality characteristics in downstream reaches of the Hanjiang River. There have been changes in the growth, spawning behaviour and overwintering condition of local fish populations, in the composition and abundance of food organisms and in the composition of the commercial fish catch. Despite the changed environment and the absence of a fish pass, fish populations are still able to grow and spawn under the new regime. Where conditions are like those of the Hanjiang River, dams may not necessarily have calamitous consequences for fishery production.

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At least three known standards are normally required for the full two-port test fixture calibration in vector network analyzers (VNA). In this paper, a calibration procedure using only one standard, based on establishing two hypothetical symmetrical fixtures using triple-through method, is shown. The results using the calibrating method to subtract the influence of fixtures are in accord with the directly measured data of the device-under-test (DUT) without the fixtures very well, which shows that the proposed method is very simple and accurate.

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The article mainly focuses on the simulation of the single electron device and circuit. The orthodox model of single electronic device is introduced and the simulation with Matlab and Pspice is illustrated in the article. Moreover, the built of robust circuit using single electronic according to neural network is done and the simulation is also included in the paper. The result shows that neural network added with proper redundancy is an available candidate for single electron device circuit. The proposed structure is also promising for the realization of low ultra-low power consumption and solution of transient device failure.

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This paper deals withmodel generation for equational theories, i.e., automatically generating (finite) models of a given set of (logical) equations. Our method of finite model generation and a tool for automatic construction of finite algebras is described. Some examples are given to show the applications of our program. We argue that, the combination of model generators and theorem provers enables us to get a better understanding of logical theories. A brief comparison between our tool and other similar tools is also presented.

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An important characteristic of virtual assembly is interaction. Traditional di-rect manipulation in virtual assembly relies on dynamic collision detection, which is very time-consuming and even impossible in desktop virtual assembly environment. Feature-matching isa critical process in harmonious virtual assembly, and is the premise of assembly constraint sens-ing. This paper puts forward an active object-based feature-matching perception mechanism and afeature-matching interactive computing process, both of which make the direct manipulation in vir-tual assembly break away from collision detection. They also help to enhance virtual environmentunderstandability of user intention and promote interaction performance. Experimental resultsshow that this perception mechanism can ensure that users achieve real-time direct manipulationin desktop virtual environment.

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In this paper, we propose a new approach to construct a 2-dimensional (2-D) directional filter bank (DFB) by cascading a 2-D nonseparable checkerboard-shaped filter pair and 2-D separable cosine modulated filter bank (CMFB). Similar to diagonal subbands in 2-D separable wavelets, most of the subbands in 2-D separable CMFBs, tensor products of two 1-D CMFBs, are poor in directional selectivity due to the fact that the frequency supports of most of the subband filters are concentrated along two different directions. To improve the directional selectivity, we propose a new DFB to realize the subband decomposition. First, a checkerboard-shaped filter pair is used to decompose an input image into two images containing different directional information of the original image. Next, a 2-D separable CMFB is applied to each of the two images for directional decomposition. The new DFB is easy in design and has merits: low redundancy ratio and fine directional-frequency tiling. As its application, the BLS-GSM algorithm for image denoising is extended to use the new DFBs. Experimental results show that the proposed DFB achieves better denoising performance than the methods using other DFBs for images of abundant textures. (C) 2008 Elsevier B.V. All rights reserved.