• Title/Summary/Keyword: Strain gradient

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Two-dimensional gel Electrophoresis of Helicobacter pylori for Proteomic Analysis

  • Jung, Tae-Sung;Kang, Seung-Chul;Choi, Yeo-Jeong;Jeon, Beong-Sam;Park, Jeong-Won;Jung, Sun-Ae;Song, Jae-Young;Choi, Sang-Haeng;Park, Seong-Gyu;Choe, Mi-Young;Lee, Byung-Sang;Byun, Eun-Young;Baik, Seung-Chul
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.97-108
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    • 2000
  • Two-dimensional gel electrophoresis (2-DE) is an essential tool of proteomics to analyse the entire set of proteins of an organism and its variation between organisms. Helicobacter pylori was tried to identify differences between strains. As the first step, whole H. pylori was lysed using high concentration urea contained lysis buffer [9.5 M Urea, 4% CHAPS, 35 mM Tris, 65 mM DTT, 0.01% SDS and 0.5% Ampholite (Bio-Rad, pH 3-10)]. The extract ($10\;{\mu}g$) was rehydrated to commercially available immobilised pH gradient (IPG) strips, then the proteins were separated according to their charges as the first dimensional separation. The IPG strips were placed on Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to separate according to molecular mass of the proteins as the second dimension. The separated protein spots were visualised by silver staining in order to compare different expression of proteins between strains. Approximately 120 spots were identified in each mini-protein electrophoresised gel, furthermore about 65 to 75 spots were regarded as identical proteins in terms of pI value and molecular weight between strains used. In addition, distinct differences were found between strains, such as 219-1, Y7 and Y14, CH150. Two representative strains were examined using strips which had pH range from 4 to 7. This strips showed a number of isoforms which were considered large spots on pH range 3-10. Furthermore, the rest of spots on pH 4-7 IPG strips appeared very distinctive compared to broad range IPG strips. 2-DE seems to be an excellent tool for analysing and identifying variations between H. pylori strains.

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Optimization of Anion-exchange Chromatography for the Separation of Agarase from Culture Broth of Pseudoalteromonas sp. (Pseudoalteromonas sp. 배양액으로부터 아가레이즈 분리를 위한 음이온교환 크로마토그래피 최적화)

  • Kim, Yu-Na;Lee, Jae-Ran;Kim, Mu-Chan;Kim, Sung-Bae;Chang, Yong-Keun;Hong, Soon-Kwang;Kim, Chang-Joon
    • Korean Chemical Engineering Research
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    • v.49 no.6
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    • pp.840-845
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    • 2011
  • Degradation products of agarose are biologically active and thus used as an ingredient in pharmaceuticals or functional cosmetics. Furthermore, it has been strongly considered as a substrate for bio-ethanol fermentation. Recently, we isolated new agarase-producing microorganism, Pseudoalteromonas sp. from south sea of Korea. In this study, we aimed to separate and purify the agarase from culture broth of this strain. Separation of agarase was performed by ion- exchange chromatography on DEAE-Sepharose resin. Equilibrium pH and volume ratio of resin to the amount of protein were optimized for the efficient adsorption of protein. 410 ${\mu}g$ of protein was completely adsorbed to 3 mL of resin at pH 7.5. The total amount of eluted protein increased as NaCl concentration increased to 400 mM at isocratic elution. Agarase was separated by linear gradient elution of NaCl (0~1,000 mM). Three major protein peaks were observed and the presence or absence of agarase in these eluted proteins was measured by Lugol's staining technique. Only six eluted protein fractions showed strong agarase activity.

Loading tests and strength evaluation of bogie frame for intermodal tram (인터모달 트램 대차프레임의 하중 시험 및 강도 평가)

  • Seo, Sung-il;Mun, Hyung-Suk;Moon, Ji-Ho;Suk, Myung-Eun;Kim, Jeong-guk
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.10
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    • pp.554-561
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    • 2016
  • In this study, loading tests and a strength evaluation of the bogie frame were conducted to verify the structural safety of the bogie system in an intermodal tram, which runs with cars on a road track. The loads were calculated taking into account the features of the road track with many sharp curves and steep gradients, which are different from the track of conventional railway. They were compared with the loads specified in the previous standard specifications. After the comparison, it was confirmed that the loads acting on the bogie system operating on a road track are slightly different from the specified loads. The specified vertical load of the standard specification for all kinds of trains is conservative, but the specified lateral and longitudinal loads are less than the calculated loads. The application of the actual loads was proven to be reasonable in the development of a new railway system. Based on the defined loads, the bogie frame was fabricated on which strain gauges were attached. It was set on the large loading frame so that the stresses could be measured when loads were applied by hydraulic actuators. After measuring the stresses, it was shown that they were below the allowable stress, which verified the structural safety of the bogie frame.

Free vibration of electro-magneto-thermo sandwich Timoshenko beam made of porous core and GPLRC

  • Safari, Mohammad;Mohammadimehr, Mehdi;Ashrafi, Hossein
    • Advances in nano research
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    • v.10 no.2
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    • pp.115-128
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    • 2021
  • In this article, free vibration behavior of electro-magneto-thermo sandwich Timoshenko beam made of porous core and Graphene Platelet Reinforced Composite (GPLRC) in a thermal environment is investigated. The governing equations of motion are derived by using the modified strain gradient theory for micro structures and Hamilton's principle. The magneto electro are under linear function along the thickness that contains magnetic and electric constant potentials and a cosine function. The effects of material length scale parameters, temperature change, various distributions of porous, different distributions of graphene platelets and thickness ratio on the natural frequency of Timoshenko beam are analyzed. The results show that an increase in aspect ratio, the temperature change, and the thickness of GPL leads to reduce the natural frequency; while vice versa for porous coefficient, volume fractions and length of GPL. Moreover, the effect of different size-dependent theories such as CT, MCST and MSGT on the natural frequency is investigated. It reveals that MSGT and CT have most and lowest values of natural frequency, respectively, because MSGT leads to increase the stiffness of micro Timoshenko sandwich beam by considering three material length scale parameters. It is seen that by increasing porosity coefficient, the natural frequency increases because both stiffness and mass matrices decreases, but the effect of reduction of mass matrix is more than stiffness matrix. Considering the piezo magneto-electric layers lead to enhance the stiffness of a micro beam, thus the natural frequency increases. It can be seen that with increasing of the value of WGPL, the stiffness of microbeam increases. As a result, the value of natural frequency enhances. It is shown that in hc/h = 0.7, the natural frequency for WGPL = 0.05 is 8% and 14% less than its for WGPL = 0.06 and WGPL = 0.07, respectively. The results show that with an increment in the length and width of GPLs, the natural frequency increases because the stiffness of micro structures enhances and vice versa for thickness of GPLs. It can be seen that the natural frequency for aGPL = 25 ㎛ and hc/h = 0.6 is 0.3% and 1% more than the one for aGPL = 5 ㎛ and aGPL = 1 ㎛, respectively.

Changes in Yeast and Bacterial Flora during Fermentation and Storage of Gugija-Liriope tuber Makgeolli using PCR-DGGE (PCR-DGGE를 이용한 구기자-맥문동 막걸리의 발효 과정과 저장 기간 중 효모와 세균 균총의 변화)

  • Min, Jin-Hong;Nam, Yun-Gyu;Ju, Jung-Il;Jung, Jae-Hong;Lee, Jong-Soo;Kim, Ha-Kun
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.111-116
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    • 2012
  • In this study, we investigated the microbial flora changes in Gugija-Liriope tuber Makgeolli during fermentation and storage periods. We brewed Gugija-Liriope tuber Makgeolli for a week through twostage fermentations and stored the fermentation broth for a month at $4^{\circ}C$ or $20^{\circ}C$. We collected the samples periodically and analyzed microbial flora changes using viable cell counts and PCR-denaturing gradient gel electrophoresis (DGGE). Yeast viable cells were seen to have decreased to 13% of pre-storage levels after storage for 15 days at $20^{\circ}C$; however significant changes were not observed during storage at $4^{\circ}C$. Prolongation of storage time dramatically decreased the availability of viable cells. Yeast viable cell numbers had decreased to 38% of pre-storage levels at $4^{\circ}C$ and 4.8% at $20^{\circ}C$ after storage for 30 days. The results of the DGGE profile for yeast showed that Saccharomyces cerevisiae and Saccharomyces sp. were the predominant strains at the beginning of fermentation and throughout the whole period of storage. Viable cell counts for total bacteria had decreased to 36% of pre-storage levels after storage for 15 days but did not significantly change for the full 30 days of storage at $4^{\circ}C$. Similarly, viable cell counts for bacteria had decreased to 5% while viable cell numbers did not significantly change for the full 30 days at $20^{\circ}C$. Viable cell counts for lactic acid bacteria were performed and the results were similar to those for total bacteria. The results of the DGGE profile for bacteria showed that Weissella cibaria was the predominant strain at the beginning of fermentation. However it had disappeared by the end of fermentation, and Lactobacillus fermentum and Pediococcus acidilactici became the predominant species during storage.