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Cotransformation of Saccharomyces cerevisiae with Heterogenous Plasmids (이종(異種) Plasmid에 의한 Saccharomyces cerevisiae의 동시형질(同時形質) 전환(轉換))

  • Kang, Byung Tae;Park, Jong Sung;Rhee, In Koo
    • Current Research on Agriculture and Life Sciences
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    • v.5
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    • pp.52-58
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    • 1987
  • The yeast S. cerevisiae DBY747 was transformed with E. C - S. C shuttle vector YIp5, YEp13 and YRp7 by the method of spheroplast. The transformation frequency of YEp13 and YRp7 in S. cerevisiae DBY747 was $1.2{\times}10^3$ and $1.0{\times}10^2$ per $10{\mu}g$ of DNA, respectively. The transformants with YIp5 plasmid incapable of autonomous replication in S. cerevisiae were not detected in the condition of this experiment, but YIpS plasmid expressed the gene carried on it when cotransformed with a helper plasmid such as YEp13 or YRp7 : autonomously replicating plasmid. When plasmids were used in covalently closed circular form, cotransformation frequency of Ylp5-YEpl3 and Ylp5-YRp7 was 210 and 95 per $10{\mu}g$ of DNA, respectively. In cotransformation of linear plasmids, transformation frequency of the same cohesive ends was similar to that of noncomplementary cohesive ends. Transformants by the cotransformation with circular plasmids have been shown much higher frequency than with linear plasmids in S. cerevisiae DBY 747. The mitotic segregation stability test suggested that the cotransformant of YIpS-YEp13 was more stable than that of YIpS-YRp7.

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Endoproteolytic Processing of Human Parathyroid Hormone in Saccharomyces cerevisiae Mutants Lacking Genes Encoding the Yap3, Mkc7, and Kex2 Endoproteases

  • Choi, Won-A.;Kang, Hyun-Ah;Chung, Bong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.679-682
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    • 1999
  • When the human parathyroid hormone (hPTH) is expressed as a secretory product in S. cerevisiae, most of the secreted hPTH is internally cleaved by endoproteolytic processing. To investigate whether the yeast endoproteases such as Kex2p, Yap3p, and Mkc7p are involved in the endoproteolytic processing of hPTH in S. cerevisiae, hPTH was expressed in S. cerevisiae mutants deficient in one or two of the following well-known endoproteases such as Kex2p, Mkc7p, and Yap3p. Among these mutants, the yap3-disrupted(yap3$\Delta$) and yap3/mkc7-disrupted (yap3Δmkc7$\Delta$) yeasts showed a significant reduction in the extent of hPTH proteolysis. In contrast, the mkc7-disrupted (mkc7$\Delta$) yeast did not reduce the proteolysis of hPTH as compared to the wild type. This suggests that Mkc7p is not involved in the endoproteolytic processing of hPTH. It was also found that the kex2-disrupted (kex2$\Delta$) mutant was not able to secrete a detectable amount of hPTH.

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The Analysis of the Change Process of Fisheries Track's High School Curriculum: Focusing on the 5th-7th Revised National Curriculum (수산계열 고등학교 교육과정의 변천과정 분석 - 제5차에서 제7차 교육과정을 중심으로 -)

  • Park, Chang-Un;Ju, Dong-Beom
    • Journal of Fisheries and Marine Sciences Education
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    • v.22 no.1
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    • pp.25-37
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    • 2010
  • This study was attempted to analyze the change process of fisheries track's high school curriculum related with the 5th-7th revised national curriculum. To accomplish this goal, the nature and goals of fisheries track's high school were reviewed. First, it was found that the fisheries track's high school was classified into three groups; high school for special goal, characterization, and industry. And the goal of fisheries track's high school was designated to harmonize the general education and vocational education in high school goal. Second, the legal basis and system of national curriculum in fisheries track's high school were discussed. The legal basis of national curriculum in fisheries track's high school was prescribed to the 'Elementary and Secondary Education Act'. The system of national curriculum was composed the general guideline and subjects. Third, the change process of fisheries track's high school national curriculum was discussed. The system of decision, general guideline, and subjects in fisheries track's high school national curriculum were seemingly to be studied on the basis of the autonomy or diversity. In conclusion, the concrete content of fisheries track's high school national curriculum was not changed compared to the 5th-7th revised curriculum.

Korea-U.S. Cross-National Comparison Study on Mathematics College Entrance Exams : the 7~th Pilot Test and the S.A.T. (한미 대학 입학 시험(수학)의 비교 연구 : 7차 실험 평가와 S.A.T.를 중심으로)

  • Kwon Oh Nam
    • The Mathematical Education
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    • v.32 no.3
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    • pp.244-255
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    • 1993
  • 본 연구를 수행하게된 동기는 1994년부터 미국에서 S.A.T.를 개정하고, 한국에서는 대학 수학 능력 시험 제도라는 새로운 제도가 도입되는 것에 있다. 대학 학업 적성 평가 제도로서 미국의 S.A.T. 제도에 대한 유효성이 많은 학자들에 의해서 연구되고 있다. 대학 수학 능력 시험과 S.A.T.는 각각 한국과 미국의 대학에서 학업 적성을 측정한다는 면에서 그 목적이 같다. 한국의 대학 수학 능력 시험의 유효성을 연구하기에는 아직 실시되지 않았으므로 너무 이르다고 본다. 대학 수학 능력 시험 제도 확립이 실험 평가에 근거하기 때문에 대학 수학 능력 시험 실험 평가와 S.A.T.를 비교 연구하는 것은 의미가 있다고 본다. 따라서 본 논문에서는 한국의 대학 수학 능력 시험 실험 평가(수리)와 미국의 S.AT.(수학)와의 상관 관계를 연구한다. 본 연구의 조사 대상으로 선발된 집단으로서 광주시의 3개교 6학급의 고등학교 3학년 283명이 참가하였다. 본 논문에서 다음과 같은 문제가 연구되었다. 1. 7차 실험 평가(수리)와 S.A.T.(수학)의 평균 점수에 대한 남녀 차이의 통계학적 유의성 (statistical significance). 2. 7차 실험 평가(수리)와 S.A.T.(수학)의 평균 점수에 대한 자연계 인문계 차이의 통계학적 유의성 (statistical significance). 3. 한국의 대학 수학 능력 시험 실험 평가(수리)와 미국의 S.A.T.(수학)의 상관 관계.

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Analysis of 2,3,7,8-substituted PCDDs, PCDFs, and DL-PCBs in muscle of crucian carp (Carassius auratus and Carassius cuvieri) from major rivers and lakes (주요 하천 및 호수에 서식하는 붕어 (Carassius auratus and Carassius cuvieri) 근육에 축적된 2,3,7,8-치환 PCDDs, PCDFs 및 DL-PCBs 분석)

  • Jeong, Gi-Ho;Moon, Ji-Yong;Moon, Dong-Ho
    • Analytical Science and Technology
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    • v.24 no.6
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    • pp.484-492
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    • 2011
  • Bioaccumulation status and distribution characteristics of PCDDs, PCDFs and DL-PCBs in cruian carp collected from the four representative sites of major river systems in Korea were investigated. The recovery rates of PCDDs and PCDFs ranged from 50.6% to 88.3%, and those of DL-PCBs ranged from 52.3% to 93.2%. The mean, median and concentration range of ${\sum}$dioxins, which represents the total concentratons of PCDDs, PCDFs and DL-PCBs accumulated in the muscle of crucian carp, were 0.39, 0.14 and 0.047-1.0 pg TEQ/g wet wt., respectively. DL-PCBs were detected above the detection limit from all the samples, whereas PCDDs and PCDFs were detected from limited crucian samples. The relative contribution of DL-PCBs to ${\sum}$dioxins was remarkably larger than those for PCDDs and PCDFs. The percent contribution was 83.6% for DL-PCBs, and followed by 12.7% and 3.7% for PCDFs and PCDDs, respectively.

20(S)-Protopanaxadiol Induces Human Breast Cancer MCF-7 Apoptosis through a Caspase-Mediated Pathway

  • Zhang, Hong;Xu, Hua-Li;Fu, Wen-Wen;Xin, Ying;Li, Mao-Wei;Wang, Shuai-Jun;Yu, Xiao-Feng;Sui, Da-Yun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7919-7923
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    • 2014
  • 20(S)-Protopanaxadiol (PPD), a ginsenoside isolated from Pananx quinquefolium L., has been shown to inhibit growth and proliferation in several cancer cell lines. The aim of this study was to evaluate its anticancer activity in human breast cancer cells. MCF-7 cells were incubated with different concentrations of 20(S)-PPD and cytotoxicity was evaluated by MTT assay. Occurrence of apoptosis was detected by DAPI and Annexin V-FITC/PI double staining. Mitochondrial membrane potential was measured with Rhodamine 123. The Bcl-2 and Bax expression were determined by Western blot analysis. Caspase activity was measured by colorimetric assay. 20(S)-PPD dose-dependently inhibited cell proliferation in MCF-7 cells, with an $IC_{50}$ value of $33.3{\mu}M$ at 24h. MCF-7 cells treated with 20(S)-PPD presented typical apoptosis, as observed by morphological analysis in cell stained with DAPI. The percentages of annexin V-FITC positive cells were 8.92%, 17.8%, 24.5% and 30.5% in MCF-7 cells treated with 0, 15, 30 and $60{\mu}M$ of 20(S)-PPD, respectively. Moreover, 20(S)-PPD could induce mitochondrial membrane potential loss, up-regulate Bax expression and down-regulate Bcl-2 expression. These events paralleled activation of caspase-9, -3 and PARP cleavage. Apoptosis induced by 20(S)-PPD was blocked by z-VAD-fmk, a pan-caspase inhibitor, suggesting induction of caspase-mediated apoptotic cell death. In conclusion, the 20(S)-PPD investigated is able to inhibit cell proliferation and to induce cancer cell death by a caspase-mediated apoptosis pathway.

Changes in Salinity, Hydraulic Conductivity and Penetration Resistance of a Silt Loam Soil in a Reclaimed Tidal Land (미사질 양토인 간척지 토양에서의 염류도와 수리전도도 및 관입 저항의 변화)

  • Jung, Yeong-Sang;Yoo, Sun-Ho;An, Yeol;Joo, Jin-Ho;Yu, Il-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.4
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    • pp.207-215
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    • 2002
  • Changes in salinity, hydraulic conductivity and penetration resistance in a reclaimed tidal land reclaimed in 1986 were studied. The salinity monitoring based on electrical conductivity of saturated extract, ECe, was conducted from 1994, when the land use for experimental crop production started after tile drainage. The site was abandoned since 1999. The hydraulic conductivity was measured by a sand fill auger hole method, and the resistance was measured with a dynamic penetrometer in situ. The averaged ECe in 1994 was $33.7dS\;m^{-1}$ ranging from 25.5 to $44.8dS\;m^{-1}$, and was decreased to $25.7dS\;m^{-1}$ with large range from 0.8 to $70.3dS\;m^{-1}$ before experiment was $1.89{\times}10^{-7}m\;s^{-1}$. It increased to $1.32{\times}10^6m\;s^{-1}$ in the top 20-cm soil with large variability, while it showed $3.44{\times}10^7m\;s^{-1}$ beneath the 20-cm soil depth with less variability. The penetration resistance of the soil ranged from 0.05 to 9.99MPa. The vertical distribution of penetration resistance indicated the hardened layer was developed at the depth of 20~40 cm where the hydraulic conductivity was sharply decreased.

Evaluation of Etiological Factors for Injuries at Oral and Maxillofacial Area (구강악안면부 외상발생의 역학적 평가)

  • Bae, Sung-Suk;Hwang, Soon-Jung
    • Journal of dental hygiene science
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    • v.12 no.4
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    • pp.310-319
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    • 2012
  • In this study, 1,495 cases of computerized data collected as disease entities of oral and maxillofacial injuries among patient visiting S university hospital located in Seoul in 2009 were analyzed and following results were obtained. It was found that the injury incidence in male was greater than in female (1.49:1), it occurred most frequently in the age of 7~14 years old (19.6%), and its most frequent cause was falling down (25.9%). Injuries occurred in other place including beach and open-air (24.7%) most frequently, often developed in anterior teeth (43.2%) and posterior teeth (43.1%), their incidence was relatively higher between 15:00~17:00 (10.4%) and 19:00~23:00 (7.1%) O'clock. In the relationship between causes of the injury and the diagnosis based on the international classification of disease), falling, fall, impingement, violence, traffic accident, sports and own making accident caused most frequently lip and oral open injuries (S01.5), open fracture of tooth (S02.51), close fracture of tooth (S02.50), close fracture of mandible (S02.60), close fracture of tooth (S02.50), respectively. From the above results, it is necessary to understand general characteristics of oral and maxillofacial injuries and to consider their trends for the establishment of systematic complement policies and for the performing safety training and public relations activities.

Synthesis and Structure-activity Relationship of Cytotoxic $5,2^I,5^I$-Trihydroxy-7,8-dimethoxyflavanone Analogues

  • Min, Byung-Sun;Ahn, Byung-Zun;Bae, Ki-Hwan
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.543-550
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    • 1996
  • Analogues of $2(S)-5, 2^{l}, 5^{l}$-trihydroxy-7, 8-dimethoxyflavanone, a naturally-occurring compound, which had been reported to have potent antitumor activity, were synthesized and examined for the cytotoxicity against three cancer cell lines. Among the intermediate chalcones and synthetic 5-hydroxy-7, 8-dimethoxyflavanone analogues, $({\pm})2^{l}, 5^{l}-dibenzyloxy-5, 7, 8-trimethoxyflavanone$ exhibited about 2-8 times stronger activity than $2(S)-5, 2^{l}, 5^{l}$-trihydroxy-7, 8-dimethoxyflavanone against L1210, K562 and A549 cancer cell lines. In the structure-activity relationship, it is suggested that among analogues of 5-hydroxy-7, 8-dimethoxyflavanone, the existence of two oxygenated groups of para-relation at $C-2^{I} and C-5^{I}$ positions on flavanone B-ring, may be necessary to exhibit effective cytotoxic activity.

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PCDDs and DFs concentrations from the fly ash and bottom ash of industrial waste incinerators in Korean (산업폐기물 소각장의 바닥재와 비산재에서 PCDDs와 DFs 농도)

  • Kim, Ki-Woong;Won, Yong Lim;Ko, Kyung Sun;Choi, Yoon Jung;Lee, Sung Kwang
    • Analytical Science and Technology
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    • v.27 no.6
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    • pp.347-351
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    • 2014
  • This study determinated the polychlorinated dibenzo-${\rho}$-dioxins and dibenzofurans (PCDDs and DFs) in fly ash and bottom ash of industrial waste incinerators. Experimental results showed that average PCDDs and DFs concentrations in fly ash were 4.5063 and 13.004 ng-TEQ/g, and in bottom ash were 0.2367 and 0.0993 ng-TEQ/g, respectively. Predominant congener in TEQ of PCDDs/DFs from fly ash were 1,2,3,4,6,7,8-HpCDD, OCDD, 2,3,4,6,7,8-HxCDF, 2,3,4,7,8-PeCDF and in bottom ash were 1,2,3,7,8-PeCDD and 2,3,4,7,8-PeCDF.