• Title/Summary/Keyword: LuxO

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Role of Alkaline Serine Protease, Asp, in Vibrio alginolyticus Virulence and Regulation of Its Expression by LuxO-LuxR Regulatory System

  • Rui, Haopeng;Liu, Qin;Wang, Qiyao;Ma, Yue;Liu, Huan;Shi, Cunbin;Zhang, Yuanxing
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.431-438
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    • 2009
  • The alkaline serine protease asp, which was shown to be a virulence factor of Vibrio alginolyticus as a purified protein, was cloned from V. alginolyticus EPGS, a strain recently isolated from moribund Epinephelus coioides in an outbreak of vibriosis in a mariculture farm of Shenzhen. The asp null mutant was constructed by homologous recombination with suicide plasmid pNQ705-1. Compared with the wild-type strain, the asp null mutant exhibited a significant decrease of total extracellular protease activity, and caused a IS-fold decrease in virulence of V. alginolyticus. In our previous study, the luxO and $luxR_{val}$ genes from V. alginolyticus MVP01 were cloned and identified, and the luxO-$luxR_{val}$ regulatory couple was shown to regulate various genes expression, suggesting that it played a central role in the quorum sensing system of V. alginolyticus. In this study, the regulation of the asp gene was analyzed by using RT-PCR and quantitative real-time PCR methods; we proved that its transcription was greatly induced at the late stage of growth and was regulated by a luxO-$luxR_{val}$ regulatory system.

Vibrio alginolyticus MviN is a LuxO-regulated Protein and Affects Cytotoxicity Towards Epithelioma Papulosum Cyprini (EPC) Cells

  • Cao, Xiaodan;Wang, Qiyao;Liu, Qin;Liu, Huan;He, Honghong;Zhang, Yuanxing
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.271-280
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    • 2010
  • Vibrio alginolyticus, a Gram-negative marine bacterium, is one of the causative agents of fish vibriosis. Its virulence factors and pathogenesis mechanism are barely known, except for some extracellular products (ECPs) that are known to be regulated by quorum sensing system. Therefore, the present study used a microarray to analyze the transcription profiles of the wild-type V. alginolyticus and a deletion mutant of luxO, the pivotal regulator in Vibrio quorum sensing systems, which resulted in the identification of a putative virulence factor, MviN. Quantitative real-time reverse transcription PCR confirmed that the transcription of mviN was upregulated in the luxO mutant when compared with wild-type, and down regulated in a luxO-con complemented strain. Furthermore, Western blotting indicated that MviN was greatly induced during the late-exponential and stationary phases of growth, indicating that the expression of MviN was cell-density dependent and quorum sensing regulated in V. alginolyticus. Meanwhile, the mviN null mutant displayed a much slower growth rate than the wild type, signifying the essential role of MviN in V. alginolyticus. Western blotting also revealed that MviN was present as an extracellular protein in V. alginolyticus. When epithelioma papulosum cyprini (EPC) cells were treated with the ECPs of the mviN mutant, no cytotoxicity was observed, whereas EPC cells treated with the wild type exhibited pathological changes, which increased with the ECPs concentration and treatment time. Therefore, the results demonstrated that MviN is a LuxO-regulated ECPs component and involved in the pathogenicity of V. alginolyticus.

Omics-Based Analysis of the luxS Mutation in a Clinical Isolate of Escherichia coli O157:H7 in Korea

  • Kim, Jong-Chul;Yoon, Jang-Won;Kim, Jong-Bae;Oh, Kyung-Hwan;Park, Mi-Sun;Lee, Bok-Kwon;Cho, Seung-Hak
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.415-424
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    • 2010
  • The purpose of this study was to investigate the relationship between the global regulatory mechanism known as quorum sensing and expression of virulence factors in Escherichia coli O157:87. A nonpolar luxS deletion was introduced into the chromosome of strain CI03J, a human clinical isolate from South Korea, to create the ${\Delta}luxS$ mutant strain ML03J. Phenotypic characterization of wild-type and mutant strains demonstrated that ML03J had no obvious growth or metabolic defects on 0.2% glucose LB medium, produced a functionally defective flagellum, and could not utilize sorbose; the biological significance of sorbose utilization is unknown. Omics-based analysis revealed the involvement of LuxS in the transcriptional activation of several flagella/chemotaxisrelated genes (flhD; fliA, C, D, S, Z; and cheA, Y, Z), repression of glutamate-dependent acid resistance genes (gadAB), and expression of virulence factors including Shiga toxin, hemolysin, and SepD within the LEE pathogenicity island.

Relationship between Environmental Conditions and the Growth of Ginseng (Panax ginseng C. A. Meyer) Plant in Field III. Field Photosynthesis under Different Light Intensity (인삼포의 환경조건과 인삼(Panax ginseng C. A. Meyer) 생육과의 관계)

  • 이성식;천성기;목성균
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.32 no.3
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    • pp.256-267
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    • 1987
  • Changes of light intensity under and above shading materials were measured at different relative light intensity(R.L.I. 5% of common straw shading, 15% of polyester shading and 20% of polyester shading at 12 o'clock on clear day) and lines(lst, 3rd and 5th lines) on clear and cloudy days in 5-year -old ginseng plant populations. Rates of photosynthesis and respiration, microclimate and root yield were also measured in field. Air temperature of R.L.I. 5% were lower 2$^{\circ}C$ compared with R.L.I. 15% and lower 3$^{\circ}C$ compared with R. L. I. 20% from 12 to 14 o'clock on clear day, but there were not difference among R. L. I. on cloudy day. Relative humidity of R. L. I. 5% were higher 5% compared compared with R.L.I. 20% from 10 to 14 o'clock on clear day, R. L. I. on cloudy day. Light intensity were below 5,000 lux at R.L.I. 5%, about 15,000 lux at R.L.I. 15% and about 20,000 lux at R.L.I. 20% from 10 to 15 o'clock on clear day. But light intensity were below 3, 000 lux at R. L. I. 5% about 10, 000 lux at R. L. I. 15% and about 15, 000 lux at R. L. I. 20% from 10 to 15 o'clock on cloudy. Photosynthetic rate of R.L.I. 15% and R.L.I. 20% were higher compared with R.L.I. 5% on clear and cloudy days. Tatal photosynthesis in a day were increased by R.L.I. 5%, 20%, 15% in turn on clear and cloudy days. R. L. I. 15% and 20% were not notable difference of photosynthetic rates among lines but R. L. I. with R. L. I. 15%, and higher 8% but there were not different among 5% was notable difference of one. Root fresh weight were increased by R.L.I. 5%, 20% and 15% in turn and R.L.I. 15% and 20% were not notable difference of root yield among lines but R. L. I. 5% was notable difference of one.

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Interaction Between the Quorum Sensing and Stringent Response Regulation Systems in the Enterohemorrhagic Escherichia coli O157:H7 EDL933 Strain

  • Oh, Kyung-Hwan;Cho, Seung-Hak
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.401-407
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    • 2014
  • Quorum sensing and the stringent response are well-known regulation systems for the expression of virulence genes in enterohemorrhagic Escherichia coli (EHEC). However, how these two systems interact is not well known. E. coli strains with mutations in two regulation systems, ${\Delta}luxS$ (ECM101) and ${\Delta}luxS{\Delta}relA{\Delta}spoT$ (ECM201), and the ${\Delta}luxS$ complement strain to ECM201 (ECM202) were created from EHEC O157:H7 EDL933 to investigate how the regulatory systems interact. The phenotypic changes of the mutant strains were characterized and compared with the wild type. The mutant strains exhibited no obvious growth defects, although acid resistance and cellular cytotoxicity were decreased significantly in all the mutant strains. Phenotypic characterization revealed that mutations in the stringent response system (ECM201 and ECM202) influenced the metabolic (defective utilization of arabinose and L-sorbose) and enzymatic activities (decreased trypsin activity, and increased ${\alpha}$-glucosidase activity). In contrast, the quorum sensing system mutant (ECM101) did not display these phenotypes. The motility of the quorum sensing system mutant (ECM101) was unchanged, but mutation in the stringent response system influenced the motility. Our results suggest that quorum sensing interacts with the stringent response regulation system.

The Light Sensitivity and Angular Dependence of the $Mg_2SiO_4$:Tb(MSO-S) TLD ($Mg_2SiO_4$:Tb(MSO-S) TLD의 광감수성과 방향의존성)

  • Kim, Do-Sung;Park, Myeong-Hwan
    • Journal of Radiation Protection and Research
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    • v.23 no.1
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    • pp.59-63
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    • 1998
  • The light sensitivity and angular dependence of the $Mg_2SiO_4$:Tb(MSO-5) TLD which affect the accuracy of the radiation dose measurement are investigated. Light-induced thermoluminescence of MSO-S TLD under the 200 lux fluorescent and the incandescent lamp for 8 hours are corresponding to 11 and 3 mR exposure, respectively. TL intensity ratio of the incident angle of ${\pm}80^{\circ}$ to normal incidence for MSO-S and badge type MSO-L are about 0.8 and 0.15, respectively.

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Effect of Light Intensity and Temperature on the Photosynthesis and Respiration of Panax spp (광도와 온도가 인삼의 광합성 및 호흡에 미치는 영향)

  • 이종화
    • Journal of Ginseng Research
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    • v.12 no.1
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    • pp.11-29
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    • 1988
  • This study was conducted to investigate the effect of light intensity and temperature on the photosynthesis and respiration of ginseng plant. Highly significant, second degree curvilinear regressions were recognized among the photosynthesis of ginseng leaves, light intensity and temperature. And an interaction between the effects of light intensity and temperature on the photosynthesis of ginseng leaves was found to be highly significant. The increasing rate of photosynthesis with the increase of light intensity was markedly decreased with increasing temperature. The light compensation point of ginseng leaves was significantly varied with temperature, and the average point was approximately 600 lux. The light saturation point of Korean ginseng was 11,000 lux at $15^{\circ}C$ and $20^{\circ}C$ and around 9,500 lux at above $25^{\circ}C$. The decreasing rate of photosynthesis with the increase of temperature significantly increased with increasing light intensity. The optimum temperature for the photosynthesis of ginseng leaves was about 15 to $22^{\circ}C$ and markedly decreased with increasing light intensity. The highest photosynthesis occurred in ginseng leaves grown with the shade of 15% transmittance. The respiration of ginseng leaves increased with the shade of 5% and/or 30% transmittance. High temperature stimulated the respiration of ginseng leaves. Percent respiration to photosynthesis of ginseng leaves grown with the shade was increased at high temperature and decreased with increasing light Intensity. It was also increased with increasing transmittance. The maximum $CO_2$ absorption of ginseng leaves grown with the shade of 5Ps and ISVS transmittance accurred at 9 o'clock a.m., whereas that of 20% transmittance occurred at 7-9 o'clock a.m. The duration of $CO_2$ absorption was distinctively long with the shade of high transmittance. The $CO_2$ compensation point in the photosynthesis of ginseng leaves was 130 ppm.

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Microbial Detection and Identification Using Biosensors

  • Kim, Sol
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.135-135
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    • 2008
  • Various biosensors were evaluated for identifying and detecting foodborne pathogens in a rapid and effective manner. First, five strains of Escherichia coli and six strains of Salmonella were identified using Fourier transform infrared spectroscopy and a statistical program. For doing this, lipopolysaccharides (LPSs) and outer membrane proteins (OMPs) were extracted from a cell wall of each bacterial strain. As a result, each strain was identifed at the level of 97% for E. coli and 100% for Salmonella. Second, E. coli O157:H7, S. Enteritidis, and Listeria monocytogenes were identified by multiplex PCR products from four specific genes of each bacteria using a capillary electrophoresis (CE). Also, ground beef for E. coli O157:H7, lettuce for S. Enteritidis, and hot dog for L. monocytogenes were used to determine the possibility of detecting pathogens in foods. Foods inoculated with respective pathogen were cultivated for six hours and multiplex PCR products were obtained and assessed. The minimum detection levels of tested bacteria were <10 cells/g, <10 cells/g, and $10^4$ cells/g for E. coli O157:H7, S. Enteritidis, and L. monocytogenes, respectively. Third, it was possible to detect S. Typhimurium in a pure culture and lettuce by a bioluminescence-based detection assay using both recombinant bacteriophage P22::luxI and a bioluminescent bioreporter. In addition, bacteriophage T4 was quantitatively monitored using E. coli including luxCDABE genes.

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Response of Rock Trout to the White Lights (백색광에 대한 쥐노래미의 반응)

  • 양용림
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.22 no.4
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    • pp.56-60
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    • 1986
  • The purpose of this study is to find the light intensity which induced maximum gathering rate and to observe the variation of the gathering rate both in daytime and at night by using rock trout, He.'~agrammos otakii (Jordan et starks). An experimental tank (360L x SOW x 55H cm) was set up in a dark room. An illumination system was attached to the end of one side of the tank to control horizontal light intensity. Six artificial light sources were prepared by combination of two light bulbs (5 W, 150 W) and seven filters. During the experiment water depth was maintained 50 em level in the tank. The tank was marked into six longitudinal sections each being 60 em long to observe the distribution of fish. The fish were acclimatized in dark condition for 50 minutes prior to the main experiment. Upon turning on the light, the number of fish in each section was counted 40 times every 30 seconds. and the gathering rate was obtained from the average number of fish in each section. The light intensity inducing maximum gathering rate was O. 7 lux (0. 5~1. 1 lux) in the daytime and 5. 2/ux (3.2-7.7 lux) at night. The variation of the gathering rate of fish in illumination time was sma II and showed the decreasing trend.

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Effects of Antioxidants on the Photosynsthesis and Carbohydrates/Saponin Contents in Panax ginseng Leaves (인삼잎의 광합성과 탄수화물.사포닌 함량에 미치는 항산화제의 효과)

  • 양덕조;김용해
    • Journal of Ginseng Research
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    • v.18 no.3
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    • pp.175-181
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    • 1994
  • We studied the folilar wiping effects of antioxidants (ascorbate, glutathione and sodium azide), which effectively inhibited the chlorophyll bleaching or completely recorved the early stage of photosynthesis of Panax ginseng C.A. Meyer, on photosynthesis, stomatal resistance, free sugar, starch, and total saponin contents of ginseng under the excess light intensity (45 kLux) during 6 days. Ascorbate and glutathione, endogenous antioxidant, recovered photosynehtsis and stomatal resistance, and reduced the photoinhibition by the excess light intensity (45 kLux) on free sugar, starch and total saponin contents. But sodium azide, exogenous $^{1}O_2$ quencher, showed negative effect. Therefore, we assumed that carbohydrates and saponin metabolisms of ginseng by antioxidants (ascorbate, glutathione) were normal. For the reduction of inhibition by excess light in ginseng a program for the higher activation of antioxidants and antioxidative enzymes in ginseng leaf will be desirable. Key words Antioxidants, ascorbate, glutathione, Photoinhibition, ginseng.

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