983 resultados para Reproducing Transformation Method


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Salinity is a major threat to sustainable agriculture worldwide. Plant NHX exchangers play an important role in conferring salt tolerance under salinity stress. In this study, a vacuolar Na+/H+ antiporter gene VrNHX1 (Genbank Accession No. JN656211.1) from mungbean (Vigna radiata) was introduced into cowpea (Vigna unguiculata) by the Agrobacterium tumefaciens-mediated transformation method. Polymerase chain reaction and Southern blot hybridization confirmed the stable integration of VrNHX1 into the cowpea genome. Comparative expression analysis by semi-quantitative RT-PCR revealed higher expression of VrNHX1 in transgenic cowpea plants than wild-type. Under salt stress conditions, T2 transgenic 35S:VrNHX1 cowpea lines exhibited higher tolerance to 200 mM NaCl treatment than wild-type. Furthermore, T2 transgenic 35S:VrNHX1 lines maintained a higher K+/Na+ ratio in the aerial parts under salt stress and accumulated higher [Na+] in roots than wild-type. Physiological analysis revealed lower levels of lipid peroxidation, hydrogen peroxide and oxygen radical production but higher levels of relative water content and proline, ascorbate and chlorophyll contents in T2 transgenic 35S:VrNHX1 lines.

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The objective of the present study was to establish a valid transformation method of Haemophilus parasuis, the causative agent of Glasser's disease in pigs, using a novel H. parasuis-Escherichia coli shuttle vector. A 4.2 kb endogenous plasmid pYC93 was extracted from an H. parasuis field isolate and completely sequenced. Analysis of pYC93 revealed a region approximately 800 bp showing high homology with the defined replication origin oriV of pLS88, a native plasmid identified in Haemophilus ducreyi. Based on the origin region of pYC93, E. coli cloning vector pBluescript SK(+) and the Tn903 derived kanamycin cassette, a shuttle vector pSHK4 was constructed by overlapping PCR strategy. When electroporation of the 15 H. parasuis serovar reference strains and one clinical isolate SH0165 with pSHK4 was performed, only one of these strains yielded transformants with an efficiency of 8.5 x 10(2) CFUhlg of DNA. Transformation efficiency was notably increased (1.3 x 10(5) CFU/mu g of DNA) with vector DNA reisolated from the homologous transformants. This demonstrated that restriction-modification systems were involved in the barrier to transformation of H. parasuis. By utilizing an in vitro DNA modification method with cell-free extracts of the host H. parasuis strains, 15 out of 16 strains were transformable. The novel shuttle vector pSHK4 and the established electrotransformation method constitute useful tools for the genetic manipulation of H. parasuis to gain a better understanding of the pathogen. (C) 2011 Elsevier B.V. All rights reserved.

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For obtaining dynamic response of structure to high frequency shock excitation spectral elements have several advantages over conventional methods. At higher frequencies transverse shear and rotary inertia have a predominant role. These are represented by the First order Shear Deformation Theory (FSDT). But not much work is reported on spectral elements with FSDT. This work presents a new spectral element based on the FSDT/Mindlin Plate Theory which is essential for wave propagation analysis of sandwich plates. Multi-transformation method is used to solve the coupled partial differential equations, i.e., Laplace transforms for temporal approximation and wavelet transforms for spatial approximation. The formulation takes into account the axial-flexure and shear coupling. The ability of the element to represent different modes of wave motion is demonstrated. Impact on the derived wave motion characteristics in the absence of the developed spectral element is discussed. The transient response using the formulated element is validated by the results obtained using Finite Element Method (FEM) which needs significant computational effort. Experimental results are provided which confirms the need to having the developed spectral element for the high frequency response of structures. (C) 2015 Elsevier Ltd. All rights reserved.

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In this paper, the transient dynamic stress intensity factor (SIF) is determined for an interface crack between two dissimilar half-infinite isotropic viscoelastic bodies under impact loading. An anti-plane step loading is assumed to act suddenly on the surface of interface crack of finite length. The stress field incurred near the crack tip is analyzed. The integral transformation method and singular integral equation approach are used to get the solution. By virtue of the integral transformation method, the viscoelastic mixed boundary problem is reduced to a set of dual integral equations of crack open displacement function in the transformation domain. The dual integral equations can be further transformed into the first kind of Cauchy-type singular integral equation (SIE) by introduction of crack dislocation density function. A piecewise continuous function approach is adopted to get the numerical solution of SIE. Finally, numerical inverse integral transformation is performed and the dynamic SIF in transformation domain is recovered to that in time domain. The dynamic SIF during a small time-interval is evaluated, and the effects of the viscoelastic material parameters on dynamic SIF are analyzed.

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In the field of fluid mechanics, free surface phenomena is one of the most important physical processes. In the present research work, the surface deformation and surface wave caused by temperature difference of sidewalls in a rectangular cavity have been investigated. The horizontal cross-section of the container is 52 mmx42 mm, and there is a silicon oil layer of height 3.5 mm in the experimental cavity. Temperature difference between the two side walls of the cavity is increased gradually, and the flow on the liquid layer will develop from stable convection to un-stable convection. An optical diagnostic system consisting of a modified Michelson interferometer and image processor has been developed for study of the surface deformation and surface wave of thermal capillary convection. The Fourier transformation method is used to interferometer fringe analysis. The quantitative results of surface deformation and surface wave have been calculated from a serial of the interference fringe patterns.The characters of surface deformation and surface wave have been obtained. They are related with temperature gradient and surface tension. Surface deformation is fluctuant with time, which shows the character of surface wave. The cycle period of the wave is 4.8 s, and the amplitudes are from 0 to 0.55 mu m. The phase of the wave near the cool side of the cavity is opposite and correlative to that near the hot side. The present experiment proves that the surface wave of thermal capillary convection exists on liquid free surface, and it is wrapped in surface deformation.

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The scattering of general SH plane wave by an interface crack between two dissimilar viscoelastic bodies is studied and the dynamic stress,intensity factor at the crack-tip is computed. The scattering problem can be decomposed into two problems: one is the reflection and refraction problem of general SH plane waves at perfect interface (with no crack); another is the scattering problem due to the existence of crack. For the first problem, the viscoelastic wave equation, displacement and stress continuity conditions across the interface are used to obtain the shear stress distribution at the interface. For the second problem, the integral transformation method is used to reduce the scattering problem into dual integral equations. Then, the dual integral equations are transformed into the Cauchy singular integral equation of first kind by introduction of the crack dislocation density function. Finally, the singular integral equation is solved by Kurtz's piecewise continuous function method. As a consequence, the crack opening displacement and dynamic stress intensity factor are obtained. At the end of the paper, a numerical example is given. The effects of incident angle, incident frequency and viscoelastic material parameters are analyzed. It is found that there is a frequency region for viscoelastic material within which the viscoelastic effects cannot be ignored.

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A novel scheme is proposed to transform a Gaussian pulse to a millimeter-wave frequency modulation pulse by using an apodized Moire fiber Bragg grating in radio-over-fiber system. The relation between the input and output pulses is analyzed theoretically by Fourier transformation method and the requirements for the proposed fiber grating are presented. An apodized Moire fiber Bragg grating is designed and its characteristics are studied. It is shown that the proposed device is feasible, and the new scheme is believed to be an effective solution for the generation of millimeter-wave sub-carrier in future radio-over-fiber systems. (c) 2006 Elsevier B.V. All rights reserved.

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The subject under investigation concerns the steady surface wave patterns created by small concentrated disturbances acting on a non-uniform flow of a heavy fluid. The initial value problem of a point disturbance in a primary flow having an arbitrary velocity distribution (U(y), 0, 0) in a direction parallel to the undisturbed free surface is formulated. A geometric optics method and the classical integral transformation method are employed as two different methods of solution for this problem. Whenever necessary, the special case of linear shear (i.e. U(y) = 1+ϵy)) is chosen for the purpose of facilitating the final integration of the solution.

The asymptotic form of the solution obtained by the method of integral transforms agrees with the leading terms of the solution obtained by geometric optics when the latter is expanded in powers of small ϵ r.

The overall effect of the shear is to confine the wave field on the downstream side of the disturbance to a region which is smaller than the wave region in the case of uniform flows. If U(y) vanishes, and changes sign at a critical plane y = ycr (e.g. ϵycr = -1 for the case of linear shear), then the boundary of this asymmetric wave field approaches this critical vertical plane. On this boundary the wave crests are all perpendicular to the x-axis, indicating that waves are reflected at this boundary.

Inside the wave field, as in the case of a point disturbance in a uniform primary flow, there exist two wave systems. The loci of constant phases (such as the crests or troughs) of these wave systems are not symmetric with respect to the x-axis. The geometric optics method and the integral transform method yield the same result of these loci for the special case of U(y) = Uo(1 + ϵy) and for large Kr (ϵr ˂˂ 1 ˂˂ Kr).

An expression for the variation of the amplitude of the waves in the wave field is obtained by the integral transform method. This is in the form of an expansion in small ϵr. The zeroth order is identical to the expression for the uniform stream case and is thus not applicable near the boundary of the wave region because it becomes infinite in that neighborhood. Throughout this investigation the viscous terms in the equations of motion are neglected, a reasonable assumption which can be justified when the wavelengths of the resulting waves are sufficiently large.

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A scheme is proposed to transform an optical pulse into a millimeter-wave frequency modulation pulse by using a weak fiber Bragg grating (FBG) in a fiber-optics system. The Fourier transformation method is used to obtain the required spectrum response function of the FBG for the Gaussian pulse, soliton pulse, and Lorenz shape pulse. On the condition of the first-order Born approximation of the weak fiber grating, the relation of the refractive index distribution and the spectrum response function of the FBG satisfies the Fourier transformation, and the corresponding refractive index distribution forms are obtained for single-frequency modulation and linear-frequency modulation millimeter-wave pulse generation. The performances of the designed fiber gratings are also studied by a numerical simulation method for a supershort pulse transmission. (c) 2007 Optical Society of America.

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A constituição de uma base de dados normativa da marcha é essencial para o diagnóstico e o tratamento de padrões atípicos da locomoção (SUTHERLAND et al., 1997). Não obstante, são escassas as informações relativas aos padrões normais da marcha de crianças (GANLEY e POWERS, 2005), carência ainda mais evidente no tocante à produção acadêmica sobre o padrão biomecânico da locomoção da população de crianças brasileiras. Nesse sentido, especialistas em análise de marcha alertam para o fato que crianças de diferentes populações podem exibir diferentes padrões de marcha de acordo com os grupos étnicos das quais foram extraídas (MORENO-HERNÁNDEZ et al., 2010). Assim sendo, o objetivo do presente estudo foi descrever o padrão biomecânico da marcha de crianças hígidas brasileiras entre 6 e 11 anos de idade. Cento e vinte e duas crianças hígidas, entre seis e 11 anos de idade foram aleatoriamente recrutadas de um universo de 328 alunos. Os sujeitos foram alocados em três grupos etários: Grupo 1 (6-7 anos), Grupo 2 (8-9 anos) e Grupo 3 (10-11 anos). Para o registro das imagens das marchas das crianças foi utilizado um sistema de captura bidimensional de movimento a uma freqüência de aquisição de 30 Hz, composto por uma câmera Sony modelo HC 46 posicionada ortogonalmente a 6 metros da pista. Marcadores esféricos reflexivos de 20mm de diâmetro foram fixados em ambos os lados do corpo dos participantes. Os valores em bruto das coordenadas dos marcadores foram transformadas em coordenadas globais 2D (CALDWELL et al., 2004) e processadas no software SkillSpector (Versão 1.0). A estratégia de Hof (1996) foi utilizada para a normalização dos dados da marcha. Os comprimentos de passo e passada apresentaram uma tendência de aumento com o avanço da idade até 8-9 anos de idade, ao passo que a cadência dos passos apresentou uma tendência de diminuição até o mesmo período. Os números não-dimensionais não apresentaram qualquer tendência de alteração com o avanço da idade. Os três grupos etários apresentaram trajetórias angulares articulares semelhantes. O presente estudo constitui ação pioneira no que tange à descrição do padrão cinemático da marcha de crianças hígidas brasileiras entre 6 e 11 anos de idade. Assim sendo, consideramos que um primeiro passo foi dado no sentido da constituição de uma base de dados normativa da locomoção desses indivíduos.

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将pBIN35S-mGFP4质粒转入受体菌DH5α中,扩繁后提取纯化质粒DNA,待棉花盛花期时,用花粉管通道转化方法将其注射到授粉24小时左右的子房中。在被检测的950个发育10~20天左右的幼胚中,发现七个幼胚在蓝光(460~490nm)激发下发出绿色荧光,而对照发出程度不同的红色荧光。将收获的“转化种子”浸泡萌发得到生长5~6天的黄化无菌苗,在200粒种子来源的无菌苗中,检出两棵转化植株。在紫外光照射灯下转化植株发出绿色荧光,其叶片和下胚轴横切面在蓝光激发下也与对照明显不同。PCR及Southern blotting结果均证实转化植株的真实性,从而为花粉管通道转化方法的可行性提供了直接可靠的细胞及分子生物学证据。 将GFPmut1质粒的gfp通过一端平接一端粘接后重组到pBI121的BamH1和Sal1限制性酶切位点从而代替GUS基因,然后将新的重组质粒用三亲交配法转入到LBA4404菌株中得到双元载体,用于棉花下胚轴切段的转化。结果表明,gfp象gus一样可作为报告基因用于农杆菌介导的棉花转化。在对筛选培养基上生长的“抗性愈伤”进一步进行报告基因检测时,只需手持紫外灯就可以检出GFP阳性愈伤,大大减少了工作量和试验费用。 另外,在进行农杆菌转化前的棉花卡那霉素敏感性实验中发现,卡那霉素对棉花下胚轴的愈伤组织形成和增殖均有明显的抑制作用,随其浓度的增加,愈伤组织形成的频率降低,增殖的倍数减小;当浓度增加到100mg/L时,愈伤组织严重褐化,其正常生长受到完全抑制;下胚轴形态学上端切段较下端更易受卡那霉素的影响。

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建立了弹性锚泊线模型,该模型根据微段垂向力学平衡关系,通过引进一个变换参数u(sinhu=dy/dx),得到了弹性悬链线的参数方程.结合工程实际,将该模型用于海洋工程中的锚泊线,给出锚泊线拉力公式,与求解算法.该算法简单有效,易于编程实现

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A transformation model for Laminaria japonica was established from 1993 to 1998, on the basis of which the transgenic kelp with heterologous gene encoding hepatitis B surface antigen (HBsAg) was obtained by using the micro-particle bombardment transformation method. Results of quantitative ELISA showed that HBsAg in transgenic kelp was 0.529 mug/mg soluble proteins on average and the highest value was 2.497 mug/mg, implying that recombinant HBsAg had natural epitope. Further support for the integration of HBsAg gene into kelp genome was obtained by PCR-Southern and total DNA hybridization. Prospect of kelp bio-reactor producing high value materials such as edible HBV vaccine was discussed as well.

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本文介绍了星球探测机器人视觉系统的标定方法,首先介绍了一种新的从视觉坐标系到车体坐标系的变换方法,随后给出了像机的模型,在像机参数优化过程中将三维重投影误差作为评价函数,利用遗传算法完成寻优过程,以保证估计出的像机参数全局最优。真实环境实验结果表明:该方法具有较高的空间定位精度。

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介绍一种利用空间向量求解坐标变换关系的方法,简化了传统的坐标系之间坐标变换关系求解的复杂计算,减小了采样误差对计算结果的影响.为建立各物体之间的位姿关系描述提供了有效的数学计算手段