• Title/Summary/Keyword: ${\alpha}시-C_2S$

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Cloning of the Cellulase Gene and Characterization of the Enzyme from a Plant Growth Promoting Rhizobacterium, Bacillus licheniformis K11 (고추역병 방제능이 있는 식물성장촉진 균주 Bacillus licheniformis K11의 cellulase 유전자의 cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.50 no.2
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    • pp.95-100
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    • 2007
  • The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

Cloning of Autoregulator Receptor Gene form Saccharopolyspora erythraea IFO 13426 (Saccharopolyspora erythraea IFO 13426으로부터 Autoregulator Receptor Protein Gene의 Cloning)

  • 김현수;이경화;조재만
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.117-123
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    • 2003
  • For screening of autoregulator receptor gene from Saccharopolyspora erythraea, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHI site of pUC19 and transformed into the E. coli DH5$\alpha$. The isolated plasmid from transformant contained the fragment of 120 bp, which was detected on 2% gel after BamHI treatment. The insert, 120 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridization using Saccha. erythraea chromosomal DNA were performed with the insert as probe. The plasmid (pEsg) having 3.2 kbp SacI DNA fragment from Saccha. erythraea is obtained. The 3.2 kbp SacI DNA fragment was sequenced by the dye terminator sequencing. The nucleotide sequence data was analyzed with GENETYX-WIN (ver 3.2) computer program and DNA database. frame analyses of the nucleotide sequence revealed a gene encoding autoregulator receptor protein which is a region including KpnI and SalI sites on 3.2 kbp SacI DNA fragment. The autoregulator receptor protein consisting of 205 amino acid was named EsgR by author. In comparison with known autoregulator receptor proteins, homology of EsgR showed above 30%.

Effect of Priming, Temperature and Light Quality on Germination of Pokeweed(Phytolacca americana) Seed (Priming, 온도 및 광질이 미국자리공 종자의 발아에 미치는 영향)

  • 강진호;류영섭;김동일;이외숙;김성희
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.2
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    • pp.153-159
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    • 1997
  • Pokeweed, a polycarpic plant, has been used as herbage medicine, vegetable or dye. It, however, is known as an aggressive plant in the vicinity of the industrial area evolving air pollutants. The experiment was done to determine the effect of priming using nitrates, germination temperature and light quality on germination of its seed to get information on the optimum germination process as well as its establishment. The daily percent germination was measured to 12 days after sowing since its seeds were treated by two different nitrates [KNO$_3$, Ca(NO$_3$)$_2$]. their different concentrations (0, 50, 150, 300mM), then treatment duration (1, 3, 6 days), different germination temperature (day /night; 30/30, 30/20, 20/30, 20/2$0^{\circ}C$) and light quality (red, white, dark) before or during germination. The percent germination was greater in the KNO$_3$ treatment than in the Ca(NO$_3$)$_2$ but in the priming treatment with KNO$_3$ in comparison with no-priming. In the priming treatment with KNO$_3$, the percent germination was increased with its increased concentration to 150mM although decreased with delayed duration to 6 days. Regardless of light quality, the greater percent germination was shown in the order of 2$0^{\circ}C$ constant and 20/3$0^{\circ}C$ alternative, 3$0^{\circ}C$ constant, 30/2$0^{\circ}C$ alternative temperature. The germination was less in the dark during germination than in the illumination in which the red light had greater percent germination compared to white light. The seeds primed with KNO$_3$ were germinated under the alternative temperature even in the dark condition.

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The Frequency of Chromosomal Aberrations of Peripheral Lymphocytes according to Radiation Dose and Dose Rate (선량 및 선량률 변화에 따른 말초혈액 임파구의 염색체 이상의 빈도)

  • Jeong Tae Sik;Baek Heum Man;Shin Byung Chul;Moon Chang Woo;Kim Mi Hyang;Lee Yong Hwan;Yum Ha Yong
    • Radiation Oncology Journal
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    • v.18 no.2
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    • pp.138-149
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    • 2000
  • Purpose : It was studied that the relationship between radiation dose, dose rate and the frequency of chromosomal aberrations in peripheral lymphocytes. Methods and Materials : Peripheral lymphocytes were irradiated in vitro with 6 MeV X-ray at dose ranges from 50 cGy to 800 cGy. The variations of the frequency of chromosomal aberrations were observed according to different radiation dose rate from 20 cGy/min to 400 cGy/min at constant total dose of 400 cGy which it was considered as factor to correct biological radiation dose measurement. Results : The yields of lymphocytes with chromosomal aberrations (dicentric chromosome, ring chromosome, acentric fragment pairs) are 0% at 50 cGy, 9% at 100 cGy, 20% at 200 cGy, 27% at 300 cGy, 55% at 400 cGy, 88% at 600 cGy, and 100% at 800 cGy. The value of Ydr is 0.000 at 50 cGy, 0.093 at 100 cGy, 0.200 at 200 cGy, 0.354 at 300 cGy, 0.612 at 400 cGy, 2.040 at 600 cGy, and 2.846 at 800 cGy. The relationship between radiation (D) and the frequency of dicentrlc chromosomes and ring Chromosomes (Ydr) can be expressed as Ydr=0.188${\times}$10$^{-2}$ D/Gy+0.422${\times}$10$^{-4}$/Gy$^{2}$${\times}$D$^{2}$ The Value of Qdr is 0.000 at 50 cGy, 1.000 at 100 cGy, 1.000 at 200 cGy, 1.333 at 300 cGy, 1.118 at 400 cGy, 2.318 at 600 cGy, and 2.846 at 800 cGy. When 400 cGy is irradiated with different dose rate each of 20, 40, 60, 80, 100, 160, 240, 320, and 400 cGy/min, Ydr is each of 0.982, 0.837, 0.860, 0.732, 0.763, 0.966, 0.909, 1.006, and 0.806, and Qdr is each of 1.839, 1.555, 1.654, 1.333, 1.381, 1.750, 1.6000, 1.710, and 1.318. Conclusion : There are not the significant variations of Ydr and Qdr values according to different dose rate. And so radiation damage is influenced by total exposed radiation doses and is influenced least of all by different dose rate when it is acute single exposure.

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Effect of Irradiation and Packaging Methods on the Oxidation of Cholesterol in Raw and Cooked Chicken Leg Meat (방사선 조사 및 포장방법이 생계육 및 조리계육의 콜레스테롤 산화에 미치는 영향)

  • Lee, J.I.;Shin, T.S.;Jin, S.K.;Kim, I.S.;Kim, Y.H.;Joo, S.T.;Park, G.B.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.825-834
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    • 2003
  • Chicken thigh from a retail market were used as experimental samples. Some chicken samples of raw state were packaged with PVDC at an aerobic and vacuum condition. The other samples were cooked until core temperature arrived at 70$^{\circ}C$ and then packaged immediately in the same way of raw samples. After samples were irradiated by electron beam at 6 kGy, they were stored in a refrigerator. Identification and quantity of cholesterol oxides were made at 0 and 7 days of storage, respectively. During the early stage of storage, 7$\beta$-hydroxycholesterol, $\alpha$,$\beta$-epoxide, cholestanetriol and 7-ketocholesterol were produced from the raw meat samples, and the production of these chemicals were significantly higher(P〈0.05) from the samples with aerobic packaging than those with vacuum packaging. With storage time, 7$\alpha$-hydroxycholesterol, 6-ketocholesterol and some other chemicals, which were not found during the early stage of storage, were found. Also, the formation of these chemicals were significantly increased(P〈0.05) with storage time. Cholesterol and lipid oxidation products of cooked meat after irradiation and irradiated meat after cooking were significantly increased(P〈0.05) with storage time for all treatments, and vacuum packaging results in showed significantly lower value(P〈0.05) than aerobic packaging. Summarizing the aforementioned results, it was found that the formation of cholesterol and lipid oxides and lipid oxidation was more easily affected by packaging condition than irradiation.

Effect on lymphocyte subpopulations of Quil A-ISCOMs with recombinant Aujeszkay's disease virus(ADV) gp50, gIII and α-ADV protein (돼지 오제스키바이러스 재조합단백질 gp50, gIII와 α-ADV을 이용한 Quil A-ISCOMs 접종시 백혈구아군 분포율에 관한 연구)

  • Moon, Jin-san;Park, Yong-ho;Jung, Suk-chan;Ku, Bok-gyeong;Lee, Sung-il;Hyun, Bang-hun;An, Soo-hwan;Davis, W.C.
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.359-369
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    • 1996
  • An effective candidate subunit vaccine was prepared by using the immunostimulating complexs(ISCOMs) with Quil A and recombinant protein(gp50, gIII and inactive $\alpha$-ADV) Aujeszky's disease virus(ADV). The weaned pigs were twice immunized with a ADV-ISCOMs, and followed by intramuscular challenge with $1{\times}10^4$ $TCID_{50}$ ADV(strain Yangsan). The unvaccinated pigs were also challenged with same dose of ADV. At 5 days after challenge, the control pigs have developed ADV clinical signs. Whereas, the vaccinated pigs protected them from ADV-induced acute symptoms and death. Also, to identify the lymphocyte subpopulation in peripheral blood with pigs from ADV-ISCOMs vaccinated and control group, lymphocyte reacted with a panel of monoclonal antibodies which are specific to swine leukocyte surface antigens and assayed by the flow cytometry. MHC class I, CD2, CD8, N cells, CD11a, and CD45 antigen positive cells were decreased after inoculating virulent ADV Yangsan strain in control group. The data indicated that ISCOMs technique was useful in ADV subunit vaccine preparation and demonstrated the importance of gp50, gIII as a component of ADV vaccine.

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Biosynthesis of the extracellular enzymes in de novo during the differentiation of Aspergillus niger (검정곰팡이의 형태분화에 따른 세포외성효소의 신생적생합성에 관한 연구)

  • Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.6 no.2
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    • pp.1-10
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    • 1978
  • In de novo biosynthesis of the extracellulor enzymes-proteinsaes, alpha and gluc-amylases during the synchronized differentiation of Aspergillus niger in submerged culture and surface liquid culture were investigated. Gluc-amylase was synthesized in the stage of presporulation in which phialide formation is involved. Proteinase was synthesized both in the stages of conidiophore formation and presporulation. Alpha-amylase was synthesized during presporulation and sporulation stages, the activity of enzyme lasted for seven days on surface liquid culture. It seemed that the synthesis was occured in de novo partly repressed by the catabolite, and its nature was found to be constitutive since it is produced in non-starch medium. Polyacrylamide gel electrophoresis have shown that presporulating and sporulating body produced diverse types of the proteins whereas the earlier stages of vegetative body showed simpler profiles. The uptake of C-14 uracil into RNA and C-14 glutamate into protein were shown to be vigorous in presporulating body rather than those in sporulating body. Coincidence of alpha-amylase biosynthesis in de novo and sporulation may be significant in the study of differentiation in which gene expression is involved.

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Fermentation Characteristics of Soybean Yogurt by Mixed Culture of Bacillus sp. and Lactic Acid Bacteria (고초균과 유산균의 혼합배양에 의한 두유 요구르트의 발효 특성)

  • Yang, Ming;Kwak, Jung Soon;Jang, Seri;Jia, Yuan;Park, Inshik
    • The Korean Journal of Food And Nutrition
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    • v.26 no.2
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    • pp.273-279
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    • 2013
  • The microorganisms producing high protease activity and acid producing ability were isolated from Chunggukjang and kimchi, which were identified as Bacillus subtilis and Lactobacillus planetarum by morphological, biochemical and nutrient requirement. The attempt was made to produce soybean milk yoghurt by using the isolated microorganisms. The mixed culture of Bacillus subtilis and Lactobacillus plantarum exhibited the lowest pH value of 4.23 and highest titratable acidity of 0.88% compared to those of single cultures at $37^{\circ}C$ for 32 hrs, and their total viable count was $4.09{\times}10^8$ $cfu/m{\ell}$. The ${\alpha}$-amylase activity was the highest in culture of Bacillus subtilis after incubation for 24 hrs, while protease activity was most produced in mixed culture of Bacillus subtilis and Lactobacillus plantarum. The amounts of reducing sugars were steadily decreased as soy milk fermentation progressed.

Effects of Acid Mine Drainage from Abandoned Coal Mines on Benthic Macroinvertebrate Communities in the Upper Reaches of the Nakdong River (낙동강 상류 폐탄광의 산성광산배수가 저서성 대형무척추동물 군집에 미치는 영향)

  • Lee, Hwang-Goo;Jung, Sang-Woo;Kim, Dong-Gun;Bae, Yeon-Jae
    • Korean Journal of Ecology and Environment
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    • v.45 no.1
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    • pp.72-81
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    • 2012
  • We investigated the effects of acid mine drainage (AMD) from abandoned coal mines on benthic macroinvertebrate communities in the upper reaches of the Nakdong River from May to October in 2009. Qualitative and quantitative sampling (Surber sampler: $50{\times}50$ cm; mesh size, 0.2 mm) was conducted at 7 study sites and 3 control sites in the study area. We thus sampled 117 species belonging to 53 families, 15 orders, 6 classes, and 5 phyla; the Ephemeroptera, Plecoptera, and Trichoptera group (EPT-group) represented the majority of the benthic macroinvertebrate community (71 species; 64.5%). In the quantitative sampling, a total of 11,575 individuals belonging to 58 species of benthic maroinvertebrates were sampled from the study sites (Sites 1-7), whereas 2,844 individuals belonging to 79 species were sampled from the control sites (Sites A-C). Tolerant species such as oligochaetes, $Epeorus$ $pellucidus$, $Baetis$ $fuscatus$, Hydropsychidae species, and Chironomidae species were predominant in the study sites. The community indices for the study sites, such as MacNaughton's dominance index (DI) (mean${\pm}$SD, $0.52{\pm}0.21$; range, 0.33-0.85) and the Shannon diversity index ($H^{\prime}$) ($2.06{\pm}0.60$; 1.06-2.57), were different from those for the control sites (DI: $0.29{\pm}0.07$, 0.22-0.35; $H^{\prime}$: $3.13{\pm}0.14$; 3.03-3.30). In the study sites, shredders and scrapers were scarce, whereas gathering-collectors (mainly Chironomidae species) were relatively abundant, as were clingers and burrowers. The detrended correspondence analysis (DCA) and similarity analyses showed that benthic macroinvertebrate communities in the study sites were clearly separated from those of the control sites, with the greatest dissimilarity being noted at the uppermost study site (Site 1), which is located close to an abandoned coal mine. The Korean saprobic index (KSI) and the ecological score using benthic macroinvertebrates (ESB) showed that the uppermost study site (Site 1) was ${\alpha}$-mesosaprobic or heavily polluted, whereas other study sites were in a fair or relatively good condition.

Studies on Selective Herbicides to Cucurbitaceae Crop (호려과작물(葫藘科作物)에 대한 선택성제초제선발(選擇性除草劑選拔)에 관한 연구(硏究))

  • Ryang, H.S.;Moon, Y.H.;Kim, N.E.;Lee, J.H.;Choi, Y.C.;Park, H.K.
    • Korean Journal of Weed Science
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    • v.8 no.2
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    • pp.208-216
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    • 1988
  • This study was conducted to select herbicides safe for cucurbitaceae crops under the polyethylene film mulching culture. No crop injury with ethalfluralin (N-ethyl-N-(2-methylally)-2, 6-dinitro-4-(trifluoromethyl) aniline) was found in gourd, water melon, cantaloup, cucumber and pumpkin of direct seeded culture. There was no significant reduction in fresh weight of gourd and pumpkin at the rate of 1080 g a.i./ha, that of water melon, cantaloup and cucumber at the rate of 720 g a.i./ha. Napropamide (N,N-diethyl-2-(${\alpha}$-naphthyloxy) propionamide) did not cause any crop injury at the rate of 1500 to 3000 g a.i./ha. There was no significant reduction in fresh weight of gourd, pumpkin and cucumber at the rate of 3000 g a.i./ha, and that of cantaloup and water melon at the rate of 1500 g a.i./ha. Trifluralin (${\alpha}$, ${\alpha}$, ${\alpha}$-trifluoro-2,6-dinitro-N, N-diprophlaniline) did not cause any crop injury in gourd. When ethalfluralin, napropamide and nitralin were applied to the transplanted seedlings of water melon and cantaloup, no significant reduction in the fresh weight were observed. The weeding effect greater than 90% was obtained with ethalfluralin at 720 to 1080 g a.i./ha and pendimethalin (N-(1-ethylpropyl)-3,4-dimethyl-2,6-dinitrobenzenamine) at 1268 g a.i./ha. The rest treatment gave the weeding effect ranging from 81 to 90%.

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