• Title/Summary/Keyword: Eco Library

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Molecular Cloning of the Gene for $\alpha$-Acylamino-$\beta$-lactam Acylhydrolase from Acetobacter turbidans by Immunochemical Detection Method (면역화학적 방법에 의한 Acetobacter turbidans의 $\alpha$-Acylamino-$\beta$-lactam Acylhydrolase의 유전자 클론화)

  • Nam, Doo-Hyun;Dewey D.Y. Ryu
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.363-368
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    • 1988
  • Molecular cloning of gene for $\alpha$-acylamino-$\beta$-lactam acylhydrolase (ALAH) III from Acetobacter turbidans has been attempted by immunochemical detection method, in which polyclonal antibody from mouse Balb/c against this enzyme was employed as a probe. As a cloning vector, λ gtll was chosen for this purpose. Two positive clones has been selected from genomic libraries of A. turbidans, which had somewhat different binding affinities on anti-ALAH III umm and anti-$\beta$-galactosidase. By restriction analysis, both clones has been turned out to lose one of EeoRI sites. From these results, it concluded that deletion of DNA between lacZ gene and inserted DNA has occurred during replication of these clones in host cells.

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Molecular cloning and restriction endonuclease mapping of homoserine dehydrogenase gene (HOM6) in yeast saccharomyces cerevisiae (Aspartate계 아미노산 대사 효모 유전자 HOM6의 cloning 및 구조분석)

  • 김응기;이호주
    • Korean Journal of Microbiology
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    • v.24 no.4
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    • pp.357-363
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    • 1986
  • Synthesis of threonine and methionine in yeast, Saccharomyces cerevisiae shares a common pathway from aspartate via homoserine. HOM6 gene encodes homoserine dehydrogenase (HSDH) which catalyzes the inter-conversion of beta-aspartate semialdehyde and homoserine. The level of HSDH is under methionine specific control. A recombinant plasmid (pEK1: 13.3kb), containing HOM6 gene, has been isolated and cloned into E. coli by complenemtary transformation of a homoserine auxotrophic yeast strain M-20-20D (hom6, trp1, ura3) to a prototrophic M20-20D/pEK1, using a library of yeast genomic DNA fragments in a yeast centromeric plasmid, YCp50(8.0kb). Isolation of HOM6has been primarily confirmed by retransformation of the original yeast strain M20-20D, using the recombinant plasmid DNA which was extracted from M20-20D/pEK1 and subsequently amplified in E. coli. Eleven cleavage sites in the insery (5.3kb) have been localized through fragment analysis for 8 restriction endonucleases; Bgl II(2 site), Bgl II(1), Cla I(3), Eco RI(1), Hind III(2), Kpn I (1), Pvu II(1) and Xho I(1).

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Isolation, Restriction Mapping, and Promoter Sequence Analysis of an Isoperoxidase Gene from Korean-Radish, Raphanus sativus L.

  • Park, Jong-Hoon;Kim, Soung-Soo
    • BMB Reports
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    • v.29 no.1
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    • pp.52-57
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    • 1996
  • A specific DNA fragment from Korean radish (Raphanus sativus L.) was amplified by performing PCR with oligonucleotide primers which correspond to the highly conserved regions of plant peroxidases. The size of the PCR product was ca. 400 bp, as expected from the known plant peroxidase genes. Comparison of the nucleotide and deduced amino acid sequences of the PCR product to those of other plant peroxidase-encoding genes revealed that the amplified fragment corresponded to the highly conserved region I and III of plant peroxidases. By screening a genomic library of Korean radish using the amplified fragment as a probe, two positive clones, named prxK1 and prxK2, were isolated. Restriction mapping studies indicated that the 5.2 kb Sail fragment of the prxK1 clone and the 4.0 kb EcoRI fragment of the prxK2 clone encode separate isoperoxidase genes. Analyses of the promoter region of the prxK1 clone shows that putative CAAT box, CMT box, and TGA1b binding sequence (5' TGACGT) are present 718 bp upstream from the start codon.

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Cloning and Expression of Pseudomonas cepacia catB Gene in Pseudomonas putida

  • Song, Seung-Yeon;Jung, Young-Hee;Lee, Myeong-Sok;Lee, Ki-Sung;Kim, Young-Soo;Kim, Chi-Kyung;Choi, Sang-Ho;Min, Kyung-Hee
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.334-340
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    • 1996
  • The enzyme, cis,cis-muconate lactonizing enzyme has been proposed to play a key role in the $\beta$-ketoadipate pathway of benzoate degradation. A 3.2-kb EcoRI fragment termed as pRSU2, isolated from a Pseudomonas cepacia genomic library was able to complement the catB defective mutant. Several relevant restriction enzyme sites were determined within the cloned fragment. In Pseudomonas putida SUC2 carrying pRSU2, the enzyme activity was relatively higher than those of the induced or partially induced state of wild type P. putida PRS2000. It was probably due to higher expression of P. cepacia catB in P. putida PRS2000. It was probably due to higher expression of P. cepacia catB in P. putida. One possible interpretation of these results is that the catB promoter in P. cepacia is recognized within P. putida, resulting in the almost same expression level.

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A Study on Differences of Contents and Tones of Arguments among Newspapers Using Text Mining Analysis (텍스트 마이닝을 활용한 신문사에 따른 내용 및 논조 차이점 분석)

  • Kam, Miah;Song, Min
    • Journal of Intelligence and Information Systems
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    • v.18 no.3
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    • pp.53-77
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    • 2012
  • This study analyses the difference of contents and tones of arguments among three Korean major newspapers, the Kyunghyang Shinmoon, the HanKyoreh, and the Dong-A Ilbo. It is commonly accepted that newspapers in Korea explicitly deliver their own tone of arguments when they talk about some sensitive issues and topics. It could be controversial if readers of newspapers read the news without being aware of the type of tones of arguments because the contents and the tones of arguments can affect readers easily. Thus it is very desirable to have a new tool that can inform the readers of what tone of argument a newspaper has. This study presents the results of clustering and classification techniques as part of text mining analysis. We focus on six main subjects such as Culture, Politics, International, Editorial-opinion, Eco-business and National issues in newspapers, and attempt to identify differences and similarities among the newspapers. The basic unit of text mining analysis is a paragraph of news articles. This study uses a keyword-network analysis tool and visualizes relationships among keywords to make it easier to see the differences. Newspaper articles were gathered from KINDS, the Korean integrated news database system. KINDS preserves news articles of the Kyunghyang Shinmun, the HanKyoreh and the Dong-A Ilbo and these are open to the public. This study used these three Korean major newspapers from KINDS. About 3,030 articles from 2008 to 2012 were used. International, national issues and politics sections were gathered with some specific issues. The International section was collected with the keyword of 'Nuclear weapon of North Korea.' The National issues section was collected with the keyword of '4-major-river.' The Politics section was collected with the keyword of 'Tonghap-Jinbo Dang.' All of the articles from April 2012 to May 2012 of Eco-business, Culture and Editorial-opinion sections were also collected. All of the collected data were handled and edited into paragraphs. We got rid of stop-words using the Lucene Korean Module. We calculated keyword co-occurrence counts from the paired co-occurrence list of keywords in a paragraph. We made a co-occurrence matrix from the list. Once the co-occurrence matrix was built, we used the Cosine coefficient matrix as input for PFNet(Pathfinder Network). In order to analyze these three newspapers and find out the significant keywords in each paper, we analyzed the list of 10 highest frequency keywords and keyword-networks of 20 highest ranking frequency keywords to closely examine the relationships and show the detailed network map among keywords. We used NodeXL software to visualize the PFNet. After drawing all the networks, we compared the results with the classification results. Classification was firstly handled to identify how the tone of argument of a newspaper is different from others. Then, to analyze tones of arguments, all the paragraphs were divided into two types of tones, Positive tone and Negative tone. To identify and classify all of the tones of paragraphs and articles we had collected, supervised learning technique was used. The Na$\ddot{i}$ve Bayesian classifier algorithm provided in the MALLET package was used to classify all the paragraphs in articles. After classification, Precision, Recall and F-value were used to evaluate the results of classification. Based on the results of this study, three subjects such as Culture, Eco-business and Politics showed some differences in contents and tones of arguments among these three newspapers. In addition, for the National issues, tones of arguments on 4-major-rivers project were different from each other. It seems three newspapers have their own specific tone of argument in those sections. And keyword-networks showed different shapes with each other in the same period in the same section. It means that frequently appeared keywords in articles are different and their contents are comprised with different keywords. And the Positive-Negative classification showed the possibility of classifying newspapers' tones of arguments compared to others. These results indicate that the approach in this study is promising to be extended as a new tool to identify the different tones of arguments of newspapers.

Change of Vegetation and Soil Characteristics of Green Roofs in Dongguk University (동국대학교 옥상녹화 지역의 식생 및 토양특성 변화)

  • Lee, Sang-Jin;Park, Gwan-Soo;Kim, Dong-Il;Lee, Dong-Kun;Kil, Sung-Ho;Jang, Seong-Wan;Park, Beom-Hwan;Yun, Jun-Young;Jang, Kwan-Woo;Lee, Ho-Young;Kwon, Oh-Jung
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.16 no.1
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    • pp.193-206
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    • 2013
  • This study was to provide the base data on the status of vegetations and soils in green roofs by analyzing the soil and vegetation characteristics of 4 green roofs in Dongguk University in September 2012. Sanglokwon(SW), Dongguk Hall(DH), University Library(UL), and Information and Culture Hall P(IC) were established in 2005, 2008, 2009, and 2010, respectively. The areas of green roofs were $700m^2$, $2,300m^2$, $1,240m^2$, and $640m^2$ in SW, DH, UL, and IC respectively. The investigated floras of vascular plants were 26 families, 55 genera, 65 species in Sanglokwon(SW), 53 families, 99 genera, 112 species in Dongguk Hall(DH), 43 families, 77 genera, 84 species in University Library(UL), and 41 families, 71 genera, 75 species in Information and Culture Hall P(IC), respectively. A positive correlation is shown between the number of plant species and planting area. Total nitrogen, organic matter, and potassium in soil have positive correlation with the number of plant species. The number of plant species was proportional to area and increased more than twice after planting. About a quarter of the invaded plants (including native and naturalized species) were naturalized plants. The total soil depths including vegetation soil and drainage soil at SW, DH, UL, and IC were 20cm, 10cm, 10cm, and 8cm, respectively. The depths of vegetation soil at SW, DH, UL, and IC were <7cm, <3cm, <2cm, and <2cm respectively. The soil pH in vegetation soil ranged from 5.22 to 5.36, and from 6.13 to 6.39 in drainage soil. Available-P concentration ranged from 10.17 to 189.77mg/kg in vegetation soil and from 6.70 to 81.17mg/kg in drainage soil. Carbon concentration in vegetation soil ranged from 2.93 to 9.70%, and 2.93 to 9.70% in drainage soil. Carbon contents in 20cm, 10cm, 10cm, and 8cm soil depths were $2.62kg/m^2$, $1.89kg/m^2$, $0.50kg/m^2$, and $0.53kg/m^2$ at SW, DH, UL, and IC, respectively.

MOLECULAR CLONING AND SEQUENCE ANALYSIS OF THE GENE FOR THE HEMIN-BINDING PROTEIN FROM Prevotella intermedia (Prevotella intermedia에서의 Hemin 결합 단백질 유전자의 분리 및 염기서열 분석)

  • Kim, Shin;Kim, Sung-Jo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.2
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    • pp.304-310
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    • 2006
  • Prevotella intermedia is one of the most frequently implicated pathogens in human periodontal disease and has a requirement for hemin for growth. This study has identified a hemin-binding P. intermedia protein by expression of a P. intermedia genomic library in Escherichia coli, a bacterium which does not require or transport exogenous hemin. The genomic library of P. intermedia was constructed into plasmid pUC18, transformed into Escherichia coli strain $DH5{\alpha}$, and screened for recombinant clones using heminbinding activity by plating onto hemin-containing agar. Approximately 5,000 recombinant E. coli colonies were screened onto LB-amp-hemin agar, single clone(pHem1) was exhibited a clearly pigmented phonotype. The 2.5kb insert DNA of pHem1 was determined by restriction enzyme mapping. Southern blot analysis of BamHI, BglII, EcoRI, HindIII and PstI-digested P. intermedia DNA indicated that single copy of the gene was present in the genome. Northern blot analysis revealed that the size of transcript was approximately 1.8 kb. The cloned gene contained a single ORF, consisting of approximately 850-residue amino acids. A BLAST search of the Institute for Genomic Research genes with similar nucleotide sequence revealed no significant similarity It needs further investigation to clarify the mechanisms of heme uptake in P. intermedia.

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Analysis of College Students' Use and Demand for Academic Information Portals: Focusing on ScienceON (대학생들의 학술정보 포털에 대한 이용 및 수요분석 - ScienceON을 중심으로 -)

  • Noh, Younghee;Wang, Dongho
    • Journal of the Korean Society for Library and Information Science
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    • v.56 no.1
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    • pp.47-65
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    • 2022
  • This study investigated college students' demand for academic information portals and conducted an improvement plan to help ScienceON become an eco-friendly portal for college students. To this end, an academic information portal similar to ScienceON was investigated and analyzed, and FGI interviews were conducted with college students to derive improvements based on the problems ScienceON felt by college students. The improvement measures proposed based on the research results are as follows. First, it is necessary to strengthen the integrated search function and instill confidence that college students can obtain all the information they want from ScienceON. Second, it is necessary to have UIs such as Google and Naver, which are preferred by college students, and improve the current hard design. Third, it is necessary to make ScienceON familiar by promoting it targeting lower grades of the university. Currently, college students belong to the MZ generation, and the MZ generation is expected to become a generation that forms a social core in the future, and related research on the MZ generation is increasing. Therefore, this study can be used as basic data for research on MZ generation, and in particular, it seems that it can be used as a reference for the demand for information search and services of MZ generation.

Molecular Cloning and Characterization of a recA-like Gene Induced by DNA Damage from a Fluorescent Pseudomonas sp.

  • Ok Bong Kim;Na Young Kim;Jae Hoon Jeong;Si Wouk Kim;Hye Gwang Jeong;Seong Myeong Yoon;Jong Kun Park;Jung Sup Lee
    • Animal cells and systems
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    • v.3 no.2
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    • pp.229-236
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    • 1999
  • The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.

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Cloning and Regulation of Schizosaccharomyces pombe Gene Encoding Ribosomal Protein S20

  • Lee, Yoon-Jong;Kim, Kyunghoon;Park, Eun-Hee;Ahn, Ki-Sup;Kim, Daemyung;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.31-36
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    • 2001
  • A cDNA clone encoding the ribosomal protein S20 has been isolated from the Schizosaccharomyces pombe cDNA library by colony hybridization. The insert contained in the original plasmid pYJ10 was transferred intro shuttle vector pRS316 generate plasmid pYJll. The dDNA insert of plasmid pYJll, contains 484 nucleotides and encodes a protein of 118 amino acids with a calculated mass of 13,544 daltons. The deduced amino acid sequence of S. pombe ribosomal protein S20 is very homologous with fruit fly, rat, and budding yeast counterparts. It is also homologous with Xenopus S22 ribosomal protein. S. pombe ribosomal protein S20 appears to be relatively hydruphobic except the C-terminal region. The 728 bp upstream region of the S20 gene was amplified from chromosomal DNA and transferred into the BamHI/EcoRI site of the promoterles $\beta$-galactosidase gene of the vector YEp357R, which resulted in fusion plasmid pYS20. The synthesis of $\beta$-galactosidase from the fusion plasmid appeared to be the highest in the mid-exponential phase. The S. pombe cells with the fusion plasmid grown at 35$\^{C}$ gave lower $\beta$-galactosidase activity than the cells grown at 30$\^{C}$. Computer analysis showed the consensus sequence CAGTCACA in the upstream regions of various ribosomal protein genes in S. pombe, which would be involved in the coordinated expression of small ribosomal proteins.

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