• Title/Summary/Keyword: 클론성

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Establishing Effective Screening Methodology for Novel Herbicide Substances from Metagenome (신규 제초활성 물질 발굴을 위한 메타게놈 스크리닝 방법 연구)

  • Lee, Boyoung;Choi, Ji Eun;Kim, Young Sook;Song, Jae Kwang;Ko, Young Kwan;Choi, Jung Sup
    • Weed & Turfgrass Science
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    • v.4 no.2
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    • pp.118-123
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    • 2015
  • Metagenomics is a powerful tool to isolate novel biocatalyst and biomolecules directly from the environmental DNA libraries. Since the metagenomics approach bypasses cultivation of microorganisms, un-cultured microorganisms that are majority of exists can be the richest reservoir for natural products discovery. To discover novel herbicidal substances from soil metagenome, we established three easy, simple and effective high throughput screening methods such as cucumber cotyledon leaf disc assay, microalgae assay and seed germination assay. Employing the methods, we isolated two active single clones (9-G1 and 9-G12) expressing herbicidal activity which whitened leaf discs, inhibited growth of microalgae and inhibited root growth of germinated Arabidopsis seeds. Spraying butanol fraction of the isolated active clones' culture broth led to growth retardation or desiccation of Digitalia sanguinalis (L) Scop. in vivo. These results represent that the screening methods established in this study are useful to screen herbicidal substances from metagenome libraries. Further identifying molecular structure of the herbicidal active substances and analyzing gene clusters encoding synthesis systems for the active substances are in progress.

Molecular Cloning of nifH, D from Frankia EuIK1 Strain, A Symbiont of Elaeagnus umbellata Root Nodules (보리수나무 뿌리혹 공생균주인 Frankia EuIK1의 nifH, D클로닝)

  • Kim, Ho-Bang;Kim, Chun-Ho;Song, Seun-Dal;An, Chung-Sun
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.258-263
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    • 1994
  • Genomic Southern hybridization of Frankia EuIKl strain, a nitrogen fixing symbiont of Elaeagnus umbellate root nodules, with nifH,D of K. pneumoniae as a probe, showed that 3.2 Kb and 5.5 Kb of BamHI fragments and 15 Kb PstI fragment were strongly hybridized with the probe, indicating nifH,D are located on these fragments. Using the same probe, one clone(pEuNIF) was isolated from the genomic library constructed into pWE15 cosmid vector by colony hybridization. The 3.2 Kb and 5.5 Kb BamHI fragments of this clone were hybridized with the same probe and this result corresponds to the genomic Southern hybridization data. However, using nifH of Frankia FaCl strain as a probe, only the 3.2 Kb BamHI fragment showed hybridization signal. Amino acid sequence deduced from nucleotide sequence of 3' terminus of the 3.2 Kb and 5' terminus of the 5.5 Kb fragments showed that the former was highly homologous with that of ArI3 nifD from 182nd to 240th amino acids, while the latter was from 241st to 282nd amino acids. These results show that nifH and partial nifD sequences are located on the 3.2 Kb fragment and residual sequences of nifH on the 5.5 Kb fragment which is contiguous to the 3.2 Kb fragment.

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Identification of Antiviral-related Genes Up-regulated in Response to Bombyx mori Nucleopolyhedrovirus (누에로부터 핵다각체병 바이러스 방어관련 유전자 정보 분석)

  • Goo, Tae-Won;Hong, Sun-Mee;Kim, Sung-Wan;Choi, Kwang-Ho;Kim, Seong-Ryul;Park, Seung-Won;Kang, Seok-Woo;Yun, Eun-Young
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.53-62
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    • 2012
  • Silkworm larvae often suffer from viral infections causing heavy losses to the economy of silk industry. Insects exhibit both humoral and cellular immune responses that are effective against various pathohens like bacteria, fungi, protozoa, etc., but no insect immune responses is effective against viral infection. To obtain genes related to insect antiviral immunity from Bombyx mori, the cDNA library was constructed from B. mori nucleopolyhedrovirus (BmNPV)-infected B. mori. From the cDNA library, we selected 411 differentially expressed clones, and the 5' ends of the inserts were sequenced to generate ESTs. In this work, 135 unigenes were generated after the assembly of 411 differentially expressed clones ESTs. Of these 135 unigenes, we selected 109 antiviral response-related candidates except 26 clones that high similarity with genes derived from BmNPV. Among 109 unigenes, a total of 80% had significant matches to genes from other organisms in the database, wheres 20% of the unigenes had not matched in the database. Functional groups of these sequences with matches in database were constructed according to their putative biological function. Three largest categories were control of cellular oraganization (52%), metabolism (20%), and protein fate (10%). The genetic information reported in this study will provide more information about antiviral-related genes in silkworms.

Production of Pyruvate Dehydrogenase Complex-E2 Specific Human Monoclonal Antibody in Fed-batch Culture Systems with High Cell Density Recombinant Escherichia coli (고농도 재조합 대장균의 Fed-batch 배양 시스템을 이용한 Pyruvate Dehydrogenase Complex-E2 특이성 인간 모노클론 항체의 생산)

  • 이미숙;전주미;차상훈;정연호
    • KSBB Journal
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    • v.15 no.5
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    • pp.489-496
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    • 2000
  • Several culture systems including batch, two-stage CSTR, semi-fed batch, and two-stage cyclic fed-batch were investigated for the efficient production of the Fab fraction of PDC-E2 specific human monoclonal antibody using high cell density recombinant E. coli. A two-phase batch system and a two-stage continuous system were examined to overcome plasmid instability problems, by separating the growth and the production stages. The cell density and productivity of the two-stage continuous culture was better than that of the two-phase batch fermentation. In the two-stage continuous culture system with DO-stat, the cell growth and the productivity were superior to those of the system without the DO control. Also, almost total plasmid stability was maintained in the two-stage continuous culture system. Modified M9 medium was selected as an optimum feeding medium for the fed-batch process, and the optimum C/N ratio determined to be 2:3. The optimum feeding rate was $0.6g/\ell/hr$ for a constant feeding strategy in semi-fed batch system. When the feeding medium was fed by pulsing, it was observed that more frequent pulsing resulted in improved cell growth. The linear feeding method was the most efficient of the various feeding methods tested. Finally, high cell density culture using a two-stage cyclic fed batch system with pH-stat was tried because the linear feeding method showed limitations in terms of obtaining high cell densities, and a cell density of $54 g/\ell$ was achieved. It was concluded that the two-stage cyclic fed batch system was the most efficient system for high cell density culture of the systems tested. However, productivity improvements were lower than expected due to the extremely high accumulations of acetate, although the low levels of residual glucose were maintained.

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The synthesis of silver powders with narrow particle distribution and spherical shape prepared by spray pyrolysis (분무열분해법에 의한 입자분포가 좁은 구형의 은 분말 제조)

  • 이교광;강윤찬;김중현;박희동
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2003.11a
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    • pp.112-112
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    • 2003
  • 은분말은 전자 산업에 있어 후막 도체 페이스트의 제조를 위해 사용되어지고 있다. 후막 페이스트는, 기재상에 스크린 프린트되고, 전도성의 회로 패턴을 형성한다. 이러한 회로는, 다음에 건조, 소성되고 액체 유기 비이클을 휘발 시키고, 그리고 은 입자를 소결시킨다. 프린트 회로 기술은 점점 고밀도이면서 더욱 정밀한 전자 회로를 요구하고 있다. 이러한 요건에 적합하기 위하여 도선은 폭이 점점 좁아지고, 선의 사이의 거리가 점점 작아지고 있다. 고밀도가 조밀하게 꽉 찬 좁은 선을 위하여 은 분말은 가능한 크기가 단일하고 구형의 형태를 가져야 한다. 현재 금속 분말을 제조하는 방법으로는 화학적 환원법, 무화 또는 분쇄, 열분해법등의 물리적 과정 및 전기 화학적 과정 등이있다. 본 연구에서는 입도 분포가 좁은 구형의 은 분말을 제조하기 위하여 기상법의 하나인 분무열분해법을 도입하였다. 또한 싸이클론을 사용하므로 큰 액적들을 걸러 입도 분포를 줄였다. 은 분말의 프리커서로써는 AgNO$_3$를 사용하였고 반응기의 온도는 $700^{\circ}C$에서 100$0^{\circ}C$까지 변화시켰으며 운반기체로써는 5%H$_2$ 혼합가스로 20L/min에서 80L/min 변화시켜 은 분말을 제조하였다. 또한 용액의 농도는 0.2M에서 1.0M까지 변화시켰다. 용액의 농도가 0.2M이고 운반기체의 유랑이 40L/min일 경우 완전한 은 상이 관찰되었고, 입자의 크기는 약 600nm였다.

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The Combustion Characteristics of a New Cyclone Jet Hybrid Combustor for Low Pollutant Emission and High Flame Stability (저공해와 고안정성을 위한 신개념의 사이클론 제트 하이브리드 연소기의 연소특성)

  • Jung, Won-Suk;Hwang, Chul-Hong;Lee, Gyou-Young;Lee, Chang-Eon
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.28 no.2
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    • pp.146-153
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    • 2004
  • A Promising new approach to achieve low pollutant emissions and improvement of flame stability is tested experimentally using a cyclone jet hybrid combustor employing both premixed and diffusion combustion mode. Three kinds of nozzle are tested for mixing enhancement of fuel and air. The LNG (Liquified Natural Gas) is used as a fuel. The combustor is operated by two methods. One is DC (Diffusion Combustion) mode generated swirl flow by air as general swirl combustor, and the other is HC (Hybrid Combustion) mode. The HC mode consists of diffusion jet flame of axial direction and premixed cyclone flame of tangential direction in order to stabilized the diffusion jet flame. The results showed that the flame stability of HC mode is significantly enhanced than that of DC mode through the change of mixing characteristics by modifications of fuel nozzle. In addition, the reductions of CO and NOx emission in HC mode, as compared with that for the DC mode, is large than about 50% in stable region. Also, even using the low calorific fuel as $CO_2$-blended gas, it is identified that the cyclone jet hybrid combustor has the high performance of flame stability.

Effects of Cloned Genes on the Stability of Shuttle Vectors between Escherichia coli and Corynebacterium glutamicum (Escherichia coli와 Corynebacterium glutamicum간의 shuttle vectors의 C. glutamicum에서의 안정성에 대한 클론된 유전자의 영향)

  • 노갑수;김성준;오종원;이현환;현형환;이재흥
    • Korean Journal of Microbiology
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    • v.29 no.3
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    • pp.149-154
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    • 1991
  • Escherichia coli/Corynebacterium glutamicum shuttle vectors, pECCG1 and pECCG2 were constructed by joining a 3.00 kb cryptic plasmid pCB 1 from C. glutamicum and a 3.94 kb plasmid pACYC 177 from E. coli. By trimming unessential parts and introducing mulitiple cloning site into the plasmid pECCG 1, a plasmid pECCG122(5.1kb) was constructed. All the shuttle vectors were stably maintained in C. glutamicum up to about 40 generations irrespective of kanamycin addition in the medium. Threonine operon (homoserine dehydrogenase/homoserine kinase) and dapA gene (dihydrodipicolinate synthetase) of C. glutamicum were cloned into the plasmid pECCG122, and the resultant plasmids were designated pTN31 and pDHDP19, respectively. They were used to study the effect of cloned foreign gene on the stability of the plasmid pECCG122. Plasmids pTN31 and pDHDP19 were segregated rapidly from C. glutamicum when cultured in the medium without kanamycin. In medium with $50\mu${\g/ml} of kanamycin, their segregation rates were much slower than those in medium without kanamycin, but the danamycin addition didn't guarantee the complete maintenance of the plasmids in C. glutamicum.

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Isolation and Elucidation of Specific RNAs by Treatment of Rhus verniciflua Stokes Extract to U937 Cell (옻추출물 처리에 의한 U937 세포에서의 특정 RNA 발현 양상)

  • Jeong, Mi-Young;Oh, Se-Wook
    • Korean Journal of Food Science and Technology
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    • v.40 no.5
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    • pp.593-598
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    • 2008
  • Differential display RT-PCR was used for screening the differentially expressed specific genes by Rhus verniciflua extract treatment to U937 cell, human leukemic monocyte. As a result, 19 clones differentially expressed were detected. Among the detected clones, one clone was confirmed to be over-expressed by R. verniciflua extract treatment in Northern blot analysis. Nucleotide sequence of the clone showed 100% homology with H2A histone family member Z gene. Therefore, it is concluded that the treatment of R. verniciflua extract to U937 cell specifically induces the expression of H2A.Z gene but its role should be elucidated by future works.

Performance of Several Jerusalem Artichoke Clones ( Helianthus tuberosus L. ) Screened for Adaptibility in Korea (돼지감자 수집클론의 우리나라 환경 적응성)

  • 임근발
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.17 no.3
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    • pp.305-314
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    • 1997
  • Nineteen clones of Jerusalem Artichoke (JA) from several countries were collected through the series of experiments about JA started in 1979. Collected clones were screened for adaptibility in Korea and showed introduction path way. The results about an ecological response of collected clones including flowering, tuberization, biomass production, sugar contents and grouping of collected clones for use of genetic material were as follows; 1. Nineteen clones collected were ffom Korea(2), Japan(l), USA(Z), Canada(2), France(4), Germany(7), and USSR(1). 2. Through the characteristics of top collected clones were divided to the types of branch-non branch, short and long plant height, and early and late maturity. Tuber characteristics were mainly grouped to the types of white skin color-violet skin color, clusters-single unit, round-elongate, and knotty-smooth. 3. Total sugar yields 6-om top at flowering time were 490 - 630kgl10a and 6-om the tuber were 420 -490 kg/ IOa through the high yielding clones. The top-high yielding clones were Mammoth French White, Fuseau 60, Nahodka, and JA3. The higher tuber yields were got from the clones of D- 19, Colombia, Bianka and Mammoth French White. 4. Collected clones were grouped to three and first group was characterized to early maturity and short plant height and second group to medium and finally, third group to late maturity and high plant height. 5. High yielding of top was 6-om the I group of early maturity and short plant height and high yielding of tuber h m III group of late maturity and high plant height.

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A Model for Optimization Process of Asbestos Dismantling Work Using Simulation (시뮬레이션을 이용한 석면 해체공사의 최적화 공정계획 모델)

  • Cho, Hyeong-Jun;Noh, Jae-Yun;Lee, Ho-Hyeon;Lee, Su-Min;Han, Seung-woo
    • Proceedings of the Korean Institute of Building Construction Conference
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    • 2022.11a
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    • pp.17-18
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    • 2022
  • In Korea, asbestos removal has been actively carried out nationwide since 2015 when asbestos was completely banned as a first-class carcinogen. Since scattering dust generated in the process of removing asbestos causes fatal diseases such as asbestos lung disease and lung cancer, concerns are growing over the safety of construction workers and building users undergoing dismantling. For this reason, regulations on asbestos sites have been strengthened and prior studies on safety and risk assessment have been conducted, but research on actual site data collection and process planning is insufficient even though safety is reduced due to delay in site construction period. Therefore, it is necessary to analyze the work and delay factors of the asbestos dismantling process and develop an optimized process plan model for workers. This study is an initial step to develop an optimized process plan model that considers the safety and productivity of asbestos dismantling work, and aims to help establish an optimized process plan for asbestos dismantling process using website clone simulation.

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