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Function of Global Regulator CodY in Bacillus thuringiensis BMB171 by Comparative Proteomic Analysis

  • Qi, Mingxia;Mei, Fei;Wang, Hui;Sun, Ming;Wang, Gejiao;Yu, Ziniu;Je, Yeonho;Li, Mingshun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.2
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    • pp.152-161
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    • 2015
  • CodY is a highly conserved protein in low G+C gram-positive bacteria that regulates genes involved in sporulation and stationary-phase adaptation. Bacillus thuringiensis is a grampositive bacterium that forms spores and parasporal crystals during the stationary phase. To our knowledge, the regulatory mechanism of CodY in B. thuringiensis is unknown. To study the function of CodY protein in B. thuringiensis, BMB171codY- was constructed in a BMB171 strain. A shuttle vector containing the ORF of cry1Ac10 was transformed into BMB171 and BMB171codY-, named BMB171cry1Ac and BMB171codY-cry1Ac, respectively. Some morphological and physiological changes of codY mutant BMB171codY-cry1Ac were observed. A comparative proteomic analysis was conducted for both BMB171codY-cry1Ac and BMB171cry1Ac through two-dimensional gel electrophoresis and MALDI-TOF-MS/MS analysis. The results showed that the proteins regulated by CodY are involved in microbial metabolism, including branched-chain amino acid metabolism, carbohydrate metabolism, fatty acid metabolism, and energy metabolism. Furthermore, we found CodY to be involved in sporulation, biosynthesis of poly-β-hydroxybutyrate, growth, genetic competence, and translation. According to the analysis of differentially expressed proteins, and physiological characterization of the codY mutant, we performed bacterial one-hybrid and electrophoretic mobility shift assay experiments and confirmed the direct regulation of genes by CodY, specifically those involved in metabolism of branched-chain amino acids, ribosomal recycling factor FRR, and the late competence protein ComER. Our data establish the foundation for in-depth study of the regulation of CodY in B. thuringiensis, and also offer a potential biocatalyst for functions of CodY in other bacteria.

The Effect of Graphene on the Electrical Properties of a Stretchable Carbon Electrode (그래핀 첨가에 따른 신축성 카본전극의 전기적 특성 변화)

  • Lee, T.W.;Park, H.H.
    • Journal of the Microelectronics and Packaging Society
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    • v.21 no.4
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    • pp.77-82
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    • 2014
  • Stretchable electrodes are focused due to many demands for soft electronics. One of the candidates, carbon black composites have advantages of low cost, easy processing and decreasing resistivity in a certain range during stretching. However, the electrical conductivity of carbon black composites is not enough for electronic devices. Graphene is 2-dimensional nanostructured carbon based material which shows good electrical properties and flexibility. They may help to improve electrical conductivity of the carbon black composites. In this study, graphene was added to a carbon black electrode to enhance electrical properties and investigated. Electrical resistivity of graphene added carbon electrode decreased comparing with that of carbon black electrode because graphene bridged non-contacting carbon black aggregates to strengthen the conductive network. Also graphene reduced an increase in the resistance of the carbon black electrode applied to strain because they connected gap of separated carbon black aggregates and aligned along the stretching direction at the same time. In conclusion, an addition of graphene to carbon black gives two benefits on the electrical properties of carbon black composite as a stretchable electrode.

A Study on Poverty Characteristics of Rural Elderly Households (농촌노인가구의 빈곤특성에 대한 비교연구 - 빈곤율과 빈곤감을 중심으로 -)

  • Kim, Young-Joo
    • Korean Journal of Social Welfare
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    • v.60 no.4
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    • pp.31-53
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    • 2008
  • This study intends to verify the poverty characteristics of the rural elderly households so as to provide more specific data for the intervention of social welfare in the improvement of quality of life of the rural elderly. According to my study of the absolute poverty ratio, the poverty gap, the relative poverty ratio, the subjective poverty ratio, the subjective economic level and the life satisfaction level of the rural elderly households in comparison with those of the other groups, the conclusion is as follows: First, the total living cost of the rural elderly households is rather low with the food cost taking about 30% of the total living cost. Second, in terms of the poverty ratio based on socio-demographic characteristics, higher poverty ratio show among women, older people and households of fewer members. Third, 82.2% of the rural elderly households living with an income below the minimum living cost is found around the poverty line. Fourth, the rural elderly households show a comparatively higher poverty ratio than the other groups in the absolute and relative poverty, but lower economic strain than other groups in the subjective poverty ratio and life satisfaction level. In short, the rural elderly households' poverty level is generally high and therefore need ways of active economic supports, while a political approach to the subjective needs of the regional and intergenerational groups is also required.

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Seismic Performance of Circular RC Columns Retrofitted Using Ductile PET Fibers (고연성 PET 섬유로 보강된 철근콘크리트 원형 기둥의 내진성능)

  • Vachirapanyakun, Sorrasak;Lim, Myung-Kwan;Choi, Dong-Uk
    • Journal of the Korea Concrete Institute
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    • v.28 no.3
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    • pp.289-298
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    • 2016
  • An experimental research was performed using fibers for the purpose of retrofitting existing reinforced concrete circular columns. Glass fiber (GF) and polyethylene terephthalate (PET) were used as well as combined GF+PET (HF). PET has high tensile strength (over 600 MPa) and high ductility (about 15%), but has very low elastic modulus (about 1/6 of GF). A total of four columns was tested against laterally applied reverse cyclic load: control column, GF-, PET-, and HF-strengthened columns. All columns retrofitted using fibers demonstrated improved moment capacity and ductility. Moment capacity of GF-, PET-, and HF-strengthened columns was 120%, 107%, and 120% of the control column, respectively. Drift ratio of all retrofitted columns also increased by 63 ~ 83% over the control column. The final failure mode of the control column was main bar buckling. The final failure mode of the GF- and HF-strengthened columns was GF rupture while that of the PET-strengthened column was main bar rupture in tension. No damage was observed for PET at the ultimate stage due to excellent strain capacity intrinsic to PET. Current test results indicate that PET can be effectively used for seismic retrofit of RC columns. It is noted that the durability characteristics of PET needs to be investigated in the future.

The Effect of Salt Contents in Diet and Water on Performance and Physiological Changes in Broiler Chicks (염분 수준이 육계 병아리의 생산성과 생리적인 변화에 미치는 영향)

  • HwangBo J.;Hong E.C.;Lee B.S.;Lee H.J.;Jo Seong-Baek;Bae H.D.;Lee S.J.;Nho W.G.
    • Korean Journal of Poultry Science
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    • v.33 no.2
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    • pp.159-164
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    • 2006
  • To investigate the effects of salt contents in diet and water on the growth of broiler chicks, 288 heads of Hubbards strain chicks, were fed with com-soybean meal diets containing 0, 0.25, 0.5, 0.75 1.0, 2.0, 3.0, or 4.0% salt from 1d to l4d posthatching. And 180 chicks were also supplemented with salt to their water at 0, 0.1, 0.2, 0.5 or 1.0%. Feed intake and weight gain tended to decrease as the salt levels in diets increased and those were the lowest in 4% group as 481 g and 168 g, however, mortality was the highest in 4% group as 75.0%. As for the salt levels in water, feed intake and weight gain were low as 427 g and 162 g in 1% group, while mortality was 77.8%. Salt poisoning chicks were shown with thirsty, appetite deficiency, rising impossibility, sleepiness, or convulsions, and in autopsy, were shown with ascites, hydropericardium, cardiac hypertrophy, hemorrhage of eyelid, hemorrhage of trachea, hemorrhage of intestine, etc. Conclusively, salt content above 4% in diet or 1% in water could reduce the performance in broiler chicks.

Rapid Detection and Discrimination of the Three Salmonella Serotypes, S. Pullorum, S. Gallinarum and S. Enteritidis by PCR-RFLP of ITS and fliC Genes

  • Cha, Se-Yeoun;Jang, Du-Hee;Kim, Sang-Min;Park, Jong-Beom;Jang, Hyung-Kwan
    • Korean Journal of Poultry Science
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    • v.35 no.1
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    • pp.9-13
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    • 2008
  • Salmonella enterica serotype gallinarum biovar Gallinarum or Pullorum and Salmonella enterica serotype Enteritidis are the most important diseases in poultry industry. Transitional diagnosis methods of these diseases such as direct isolation and identification by a biochemical test are time consuming with low specificity. In this study, we have focused on the suitable procedure for the rapid and accurate diagnosis of diseases derived from the three Salmonella strains. We initially confirmed Salmonella species by PCR using a specific ITSF/ITSR primer pair instead of biochemical test, and then the PCR-amplified phase 1 flagellin (fliC) using a specific fliCF/fliCR primer pair was digested with a restriction endonuclease, Bpm I and/or Bfa I, to discriminate among S. Pullorum, S. Gallinarum, and S. Enteritidis. We found that these methods could be applied to field isolates of the three Salmonella strains to detect and to discriminate rapidly for convenient diagnosis.

Comparative Study of Toxicological Methods and Field Resistance to Insecticides in Diamondback moth(Lepidoptera: Plutellidae) (배추좀나방(Plutella xylostella L.)의 독성시험방법 비교와 지역별 약제저항성에 관한 연구)

  • 이승찬;조영식;김도익
    • Korean journal of applied entomology
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    • v.32 no.3
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    • pp.323-329
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    • 1993
  • These studies were conducted to evaluate the five comparative test methods for detecting chemical resistance and to investigate resistant level of field populations of diamondback moth (Plutella xylostella L.). Leaf disc method was practically rocomrnendable because of its rapidity and low CV(l1.4%). Topical application method was a precise replicabiliLy(CV=8.00/0) but it was time consuming and difficult in mampulation. The other 3 methods showed higher CV ranging from 14.9% to 21.4%. Based on $LC_{50}$ values by topical application method, field populations of diamondback moth collected from 4 different regions, Kwangju, Kimhae, Jeju, and Inje to prothiofos showed from 3.3 to 61.1 times higher resistance than the susceptible strain, whereas to cypermethrin, Lhey were from 7.5 to 141.7 times higher than the susceptible. To cartap hydrochloride, they showed from 10.5- to 33.3-fold resistant levels as high as the susceptible. Finally, based on $LC_{50}$ values to Bacillus thuringiensis by leaf disc technique, the resistant levels of the field populations were from 1.9 Lo 8.1 times as compared to the susceptible.

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Physiological Characteristics and GABA Production of Lactobacillus plantarum K74 isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum K74의 GABA 생산 및 생리적 특성)

  • Park, Sun-Young;Shim, Hye-Young;Kim, Kee-Sung;Lim, Sang-Dong
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.143-152
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    • 2013
  • Aminobutyric acid (GABA), a major inhibitory neurotransmitter in the central nervous system of animals, has several physiological effects including anti-hypertensive, diuretic, tranquilizing, and anti-stress properties, in humans. The purpose of this study was to investigate Lactobacillus plantarum K74, which was isolated from kimchi and selected as a strain with a high ability to produce GABA, to develop a new starter culture for fermented milk production. L. plantarum K74 produced $134.52{\mu}g/mL$ GABA in MRS broth containing 1% MSG, $212.27{\mu}g/mL$ GABA in MRS broth containing 2% MSG, and $234.63{\mu}g/mL$ GABA in MRS broth containing 3% MSG. The optimum growth temperature of L. plantarum K74 was $34^{\circ}C$, reaching a pH of 4.4 after 18 hours of growth. L. plantarum K74 was most sensitive to novobiocin out of 16 different antibiotics tested, and was most resistant to kanamycin and polymyxin B. L. plantarum K74 did not produce ${\beta}$-glucuronidase, a carcinogenic enzyme, and was comparatively tolerant to bile juice and low pH. Furthermore, it displayed resistance to Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus at rates of 54.9%, 46.3%, and 0.7%, respectively.

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Genetic Diversity, Pathogenicity, and Fungicide Response of Fusarium oxysporum f. sp. fragariae Isolated from Strawberry Plants in Korea (국내 딸기 시들음병균 Fusarium oxysporum f. sp. fragariae의 유전적 다양성, 병원성과 살균제 반응)

  • Nam, Myeong Hyeon;Kim, Hyun Sook;Park, Myung Soo;Min, Ji Young;Kim, Heung Tae
    • Research in Plant Disease
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    • v.26 no.2
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    • pp.79-87
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    • 2020
  • Fusarium wilt caused by Fusarium oxysporum f. sp. fragariae (Fof) is the most important diseases of a strawberry field in Korea. We surveyed phylogenetic analysis, pathogenicity test, and fungicide response about Fof isolates isolated from Korea. Twenty-seven isolates of F. oxysporum isolated from strawberry plants were conducted in this study. Specific amplification by Fof specific primer was confirmed in all 26 isolates except Fo080701 isolate. The nuclear ribosomal intergenic spacer region and the translation elongation factor EF-lα gene sequences of isolates revealed three main lineages. Most of all isolates were contained DNA lineage group 1, but 2 and 3 group was shown only one and three isolates, respectively. All isolates were shown in pathogenicity with cv. Seolhyang. The EC50 mean values of prochloraz ranged 0.02-0.1 ㎍/ml except for Fo080701 and effectively inhibited mycelial growth at low concentrations. The EC50 value of metconazole was also 0.04-0.22 ㎍/ml, showing a similar inhibitory effect to that of prochloraz. The EC50 value of pyraclostrobin was 0.23-168.01 ㎍/ml, which was different according to the strain. In the field trial, boscalid+fludioxonil, fluxapyroxad+pyraclostrobin, and prochloraz manganese were selected as the effective fungicides for controlling Fusarium wilt.

Effects of Selenium and Zinc on the Toxicity of Cadmium in Rat (백서에서 셀레늄 및 아연이 카드뮴 독성에 미치는 영향)

  • 강영미;이용욱
    • Journal of Environmental Health Sciences
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    • v.16 no.1
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    • pp.29-43
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    • 1990
  • The purpose of this study is to investigate the vairous change in the toxicity of cadmium by the simultaneous administration of selenium and zinc, which have been reported to change -the toxicity of cadmium through the interaction with cadmium, to rat. For the experiment, 42 rats of Sprague-Dawley strain were used. The experimental groups were divided into 6 groups: a control group, a cadmium (100ppm) alone treatment group, a cadmium (100ppm) and zinc (100ppm) combined treatment group, and three cadmium (100ppm), zinc (100ppm) and selenium (1, 4, and 8ppm) combined treatment groups. The rats were allocated seven to each group and observed for seven weeks. The results of experiment are as follows: 1. The food consumptions of each group were reduced, compared with a control group, especially, in a cadmium and zinc combined treatment group and a cadmium $\cdot$ zinc and selenium(1ppm) combined treatment group to the significant level compared with a control group (p < 0.05). The water consumptions of each group were reduced to the very significant level compared with a control group (p < 0.01). The feed efficiencies of each group were lower than a control group, and among them the highest group was cadmium $\cdot$ zinc and selenium (8ppm) combined treatment group as 90% of a control group. 2. In all groups, the weight gains were highest in the second week and the total weight gains were reduced to the very significant level compared with a control group (p < 0.01). 3. In all groups, the relative weights of liver were reduced, compared with a control group, especially, a cadmium alone treatment group was reduced to the significant level (p < 0.05). The relative weight of kidney was high to the significant level in a cadmium alone treatment group (p < 0.05) compared with a control group. In all groups, the relative weights of testis were reduced, compared with a control group, but the levels were not significant. 4. The accumulation of cadmium was highest in the kidney and the order of height was in liver, testis and blood, respectively. In all groups, the amount of cadmium accumulation was high to the very significant level compared with a control group (p < 0.01). In liver, the amount of acdmium accumulation in. a cadmium alone treatment group was high to the significant level compared with a cadmium $\cdot$ zinc and selenium (8ppm) combined treatment group (p < 0.05), and in kidney, the amount of cadmium accumulation in a cadmium alone tretment group was high to the very significant level compared with the cadmium $\cdot$ zinc and selenium (4, 8ppm) combined treatment groups (p < 0.01). However, in testis, among the treatment groups the level was not significant and in blood, a cadmium alone treatment group was low to the significant level compared with the cadmium $\cdot$ zinc and selenium (4, 8ppm) combined treatment groups (p < 0.05). 5. According to the histopathological finding on the testis, some of the seminiferous tubules of a group treated with cadmium alone showed severe necrosis and atrophy. But the testis of cadmium $\cdot$ zinc and selenium (8ppm) combined treatment group was similar to that of a control group. From the results of this study, it can be concluded that the repeated simultaneous oral administration of large doses of selenium with the cadmium produces the partial amelioration of cadmium toxicity, whereas zinc does not.

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