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Characterization of the nar Promoter of Escherichia coli to use as an inducible promoter in Wild-type host Agrobacterium.tumefaciens

  • Lee, Gil-Ho;Jo, Mu-Hwan;Lee, Jong-Won
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.758-761
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    • 2001
  • In this study, the nar promoter of E. coli was characterized to see whether the nar promoter cloned onto pBBR122 can be used as an expression promoter of gram negative microbes. For this purpose, a plasmid with lacZ gene expressing ${\beta}-galactosidase$ instead of the structural genes of nar operon in a gram negative host strain(Agrobacterium.tumefaciens) was used to simplify an assay of induction of the nar promoter. The following effects were investigated to find optimal conditions: methods of inducing the nar promoter, optimal nitrate concentration, maximally inducing the nar promoter, the amount of expressed ${\beta}-galactosidase$ and induction ratio(specific ${\beta}-galactosidase$ activity after maximal induction/specific ${\beta}-galactosidase$ activity before induction). The following results were obtained from the experiments: the growth of Agrobacterium with E.coli nar promoter was not much affected by nitrate concentration in the shake-flask; induction of nar promoter was optimal when Agrobacterium was grown in the presence of 1% nitrate ion at the beginning of culture and when overnight culture was completely grown in the shake-flask before being transferred to other shake-flask; the amount of ${\beta}-galactosidase$ per cell and per medium volume was maximal when Agrobacterium was grown under aerobic condition to $OD_{600}$ of 1.7; then the nar promoter was induced under microaerobic and anaerobic condition made by lowering dissolved oxygen level(DO). After 2-3h of induction in the YEP medium selected as a main culture medium, the specific ${\beta}-galactosidase$ activity became about 17,000 Miller units in the fermentor cluture.

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Optimization of Submerged Culture Conditions for Mycelial Growth and Exopolysaccharides Production by Agaricus blazei

  • Kim, Hyun-Han;Na, Jeong-Geol;Chang, Yong-Keun;Chun, Gie-Taek;Lee, Sang-Jong;Jeong, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.944-951
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    • 2004
  • The influences of inoculum size, pH, and medium composition on mycelial growth and exopolysaccharides (EPS) production were investigated in shake flasks and in a bioreactor. The optimum inoculum size for both mycelial growth and EPS production was identified to be 10% (v/v) in shake flask cultures. The optimal initial pH for mycelial growth and EPS production in shake flask cultures were found to be 5.0 and 7.0, respectively. However, the optimal pH was 5.0 for both mycelial growth and EPS production in bioreactor cultures where the pH was regulated. The optimal mass ratio of the two major carbon sources, glucose to dextrin, was 1:4. The optimal mass ratio of the two major nitrogen sources, yeast extract to soy tone peptone, was 2:1. When 500 mg $1^{-1}$ of $MnSO_4-5H_2O$ was added to the bioreactor culture, both mycelial growth and EPS production were enhanced by approximately 10%. Under the optimized conditions, a mycelial biomass of 9.85 g $1^{-1}$ and an EPS concentration of 4.92 g $1^{-1}$ were obtained in 4 days.

Development of Miniaturized Culture Systems for Large Screening of Mycelial Fungal Cells of Aspergillus terreus Producing Itaconic Acid

  • Shin, Woo-Shik;Lee, Dohoon;Kim, Sangyong;Jeong, Yong-Seob;Chun, Gie-Taek
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.101-111
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    • 2017
  • The task of improving a fungal strain is highly time-consuming due to the requirement of a large number of flasks in order to obtain a library with enough diversity. In addition, fermentations (particularly those for fungal cells) are typically performed in high-volume (100-250 ml) shake-flasks. In this study, for large and rapid screening of itaconic acid (IA) high-yielding mutants of Aspergillus terreus, a miniaturized culture method was developed using 12-well and 24-well microtiter plates (MTPs, working volume = 1-2 ml). These miniaturized MTP fermentations were successful, only when highly filamentous forms were induced in the growth cultures. Under these conditions, loose-pelleted morphologies of optimum sizes (less than 0.5 mm in diameter) were casually induced in the MTP production cultures, which turned out to be the prerequisite for the active IA biosynthesis by the mutated strains in the miniaturized fermentations. Another crucial factor for successful MTP fermentation was to supply an optimal amount of dissolved oxygen into the fermentation broth through increasing the agitation speed (240 rpm) and reducing the working volume (1 ml) of each 24-well microtiter plate. Notably, almost identical fermentation physiologies resulted in the 250 ml shake-flasks, as well as in the 12-well and 24-well MTP cultures conducted under the respective optimum conditions, as expressed in terms of the distribution of IA productivity of each mutant. These results reveal that MTP cultures could be considered as viable alternatives for the labor-intensive shake-flask fermentations even for filamentous fungal cells, leading to the rapid development of IA high-yield mutant strains.

An Efficient Vehicle Image Compensation Algorithm based on Histogram Equalization (히스토그램 균등화 기반의 효율적인 차량용 영상 보정 알고리즘)

  • Hong, Sung-Il;Lin, Chi-Ho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.3
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    • pp.2192-2200
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    • 2015
  • In this paper, we propose an efficient vehicle image compensation algorithm based on Histogram Equalization. The proposed a vehicle image compensation algorithm was elimination to the vehicle image shake using motion compensation and motion estimation. And, algorithm was calculated the histogram of pixel values from each sub-image by dividing the image as the constant size areas in order to image enhancement. Also, it had enhancement to the image by adjusting the gradient. The proposed algorithm was evaluate the difference between of performance and time, image by applied to the IP, and were confirmed the image enhancement with removing of vehicle camera image shake. In this paper, the proposed vehicle image enhancement algorithm was demonstrated effectiveness when compared to existing vehicle image stabilization, because the elimination of shake for the vehicle images used real-time processing without using a memory. And it was obtained the reduction effect of the computation time by the calculated through block matching, and obtained the better restoration result for naturalness of the image with the lowest noise.

A Study on the Evaluation of Soil Nonlinear Characteristics by Seismic Recorded Data at Downhole Array (Downhole 지진계측자료에 의한 지반의 비선형성 평가에 관한 연구)

  • 장정범;서용표;이종림;이계희
    • Journal of the Earthquake Engineering Society of Korea
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    • v.5 no.6
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    • pp.29-35
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    • 2001
  • The soil-structure interaction(SSI) analysis is essential to soil site where shear wave velocity is less than 1,050 m/sec and soil nonlinear characteristics for this kind of soil site have to be considered in SSI analysis. In order to consider soil nonlinear characteristics in the SSI analysis, simple and reliable soil nonlinear evaluation technique with seismic recorded data at downhole array is proposed in this study. The SSI analysis is carried out in order to prove the reliability of the proposed evaluation technique with Hualien large scale seismic test(HLSST) site in Taiwan. The analytical result are compared with Hualien earthquake recorded data and the analytical results with SHAKE program which is prevailed at present. As a result, the proposed evaluation technique shows a good agreement with both the Hualien earthquake recorded data and the analytical result with SHAKE program and the reliability and usefulness are confirmed.

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Shake-table study of plaster effects on the behavior of masonry-infilled steel frames

  • Baloevic, Goran;Radnic, Jure;Grgic, Nikola;Matesan, Domagoj
    • Steel and Composite Structures
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    • v.23 no.2
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    • pp.195-204
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    • 2017
  • The effects of plaster on the behavior of single-story single-bay masonry-infilled steel frames under in-plane base accelerations have been experimentally investigated by a shake-table. Tested structures were made in a 1/3 scale, with realistic material properties and construction methods. Steel frames with high and low flexural rigidity of beams and columns were considered. Each type of frame was tested with three variants of masonry: (i) non-plastered masonry; (ii) masonry infill with conventional plaster on both sides; and (iii) masonry infill with a polyvinyl chloride (PVC) net reinforced plaster on both sides. Masonry bricks were made of lightweight cellular concrete. Each frame was firstly successively exposed to horizontal base accelerations of an artificial accelerogram, and afterwards, to horizontal base accelerations of a real earthquake. Characteristic displacements, strains and cracks in the masonry were established for each applied excitation. It has been concluded that plaster strengthens the infill and prevents damages in it, which results in more favorable behavior and increased bearing capacity of plastered masonry-infilled frames compared to non-plastered masonry-infilled frames. The load-bearing contribution of the adopted PVC net in the plaster was not noticeable for the tested specimens, probably due to relative small cross section area of fibers in the net. Behavior of masonry-infilled steel frames significantly depends on frame stiffness. Strong frames have smaller displacements than weak frames, which reduces deformations and damages of an infill.

Earthquake Response Analysis for Three-Story Building with Reinforced Concrete Shear Walls (3층 철근콘크리트 전단벽 구조물의 지진응답해석)

  • Rhee, Inkyu;Lee, Eun-Haeng;Kim, Jae-Min
    • Journal of the Earthquake Engineering Society of Korea
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    • v.25 no.3
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    • pp.103-110
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    • 2021
  • A shake table test is conducted for the three-story reinforced concrete building structure using 0.28 g, 0.5 g, 0.75 g, and 1.0 g of seismic input motions based on the Gyeongju earthquake. Computational efforts are made in parallel to explore the mechanical details in the structure. For engineering practice, the elastic modulus of concrete and rebar in the dynamic analysis is reduced to 38% and 50%, respectively, to calibrate the structure's natural frequencies. The engineering approach to the reduced modulus of elasticity is believed to be due to the inability to specify the flexibility of the actual boundary conditions. This aspect may lead to disadvantages of nonlinear dynamic analysis that can distort local stress and strain relationships. The initial elastic modulus can be applied directly without the so-called engineering adjustment with infinite element models with spring and spring-dashpot boundary conditions. This has the advantage of imposing the system flexibility of the structure on the sub-boundary conditions of springs and damping devices to control its sensitivity in a serial arrangement. This can reflect the flexibility of realistic boundary conditions and the effects of system damping (such as the gap between a concrete footing and shake table, loosening of steel anchors, etc.) in scalar quantities. However, these spring and dashpot coefficients can only be coordinated based on experimental results, making it challenging to select the coefficients in-prior to perform an experimental test.

Use of large-scale shake table tests to assess the seismic response of a tunnel embedded in compacted sand

  • Zhou, Hao;Qin, Xiaoyang;Wang, Xinghua;Liang, Yan
    • Earthquakes and Structures
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    • v.15 no.6
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    • pp.655-665
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    • 2018
  • Shield tunnels are widely used throughout the world. However, their seismic performance has not been well studied. This paper focuses on the seismic response of a large scale model tunnel in compacted sand. A 9.3 m long, 3.7 m wide and 2.5 m high rigid box was filled with sand so as to simulate the sandy soil surrounding the tunnel. The setup was excited on a large-scale shake table. The model tunnel used was a 1:8 scaled model with a cross-sectional diameter of 900 mm. The effective shock absorbing layer (SAL) on the seismic response of the model tunnel was also investigated. The thickness of the tunnel lining is 60 mm. The earthquake motion recorded from the Kobe earthquake waves was used. The ground motions were scaled to have the same peak accelerations. A total of three peak accelerations were considered (i.e., 0.1 g, 0.2 g and 0.4 g). During the tests, the strain, acceleration and soil pressure on the surface of the tunnel were measured. In order to investigate the effect of shock absorbing layer on the dynamic response of the sand- tunnel system, two tunnel models were set up, one with and one without the shock absorbing layer of foam board were used. The results shows the longitudinal direction acceleration of the model tunnel with a shock absorbing layer were lower than those of model tunnel without the shock absorbing layer, Which indicates that the shock absorbing layer has a beneficial effect on the acceleration reduction. In addition, the shock absorbing layer has influence on the hoop strain and earth pressure of the model tunnel, this the effect of shock absorbing layer to the model tunnel will be discussed in the paper.

Influence of multi-component ground motions on seismic responses of long-span transmission tower-line system: An experimental study

  • Tian, Li;Ma, Ruisheng;Qiu, Canxing;Xin, Aiqiang;Pan, Haiyang;Guo, Wei
    • Earthquakes and Structures
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    • v.15 no.6
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    • pp.583-593
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    • 2018
  • Seismic performance is particularly important for life-line structures, especially for long-span transmission tower line system subjected to multi-component ground motions. However, the influence of multi-component seismic loads and the coupling effect between supporting towers and transmission lines are not taken into consideration in the current seismic design specifications. In this research, shake table tests are conducted to investigate the performance of long-span transmission tower-line system under multi-component seismic excitations. For reproducing the genuine structural responses, the reduced-scale experimental model of the prototype is designed and constructed based on the Buckingham's theorem. And three commonly used seismic records are selected as the input ground motions according to the site soil condition of supporting towers. In order to compare the experimental results, the dynamic responses of transmission tower-line system subjected to single-component and two-component ground motions are also studied using shake table tests. Furthermore, an empirical model is proposed to evaluate the acceleration and member stress responses of transmission tower-line system subjected to multi-component ground motions. The results demonstrate that the ground motions with multi-components can amplify the dynamic response of transmission tower-line system, and transmission lines have a significant influence on the structural response and should not be neglected in seismic analysis. The experimental results can provide a reference for the seismic design and analysis of long-span transmission tower-line system subjected to multi-component ground motions.

Microbial Desulfurization of a Bituminous Coal by Iron-Oxidizing Bacteria Thiobacillus ferooxidans (철산화 박테리아 Thiobacillus ferrooxidans에 의한 역청탄의 생물학적 탈황)

  • 류희욱
    • KSBB Journal
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    • v.11 no.2
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    • pp.238-245
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    • 1996
  • Microbial desulfurization characterlstics of a bituminous coal have been determined by using Thiobacillus ferrooxidans. The effects of process variables (such as coal pulp density, particle size and addition of surfactants) on pyrite removal have been investigated in both shake and airlift-bioreactor culture experiments. In shake experiments, pyrite could be removed over 78% for pulp densifies below 20% (w/v) and removed below 40% for pulp densities over 30% (w/v) in 8 days. Pyrite removal decreased with increasing pulp densities, and it also decreased sharply with increasing particle sizes. In airlift bioreactor experiments, pyrite at 50% (w/v) pulp density could be removed about 50%. Its value is much higher than 15% at the same pulp density in a shake experiment. With addition of surfactants, pyrite removal was enhanced in shake experiments significantly, whereas it was slightly decreased in an airlift bioreactor experiment.

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