• 제목/요약/키워드: producing

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A Proteomic Approach to Study msDNA Function in Escherichia coli

  • Jeong, Mi-Ae;Lim, Dongbin
    • Journal of Microbiology
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    • 제42권3호
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    • pp.200-204
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    • 2004
  • Retron is a prokaryotic genetic element that produces multicopy single-stranded DNA covalently linked to RNA (msDNA) by a reverse transcriptase. It was found that cells producing a large amount of msDNA, rather than those that did not, showed a higher rate of mutation. In order to understand the molecular mechanism connecting msDNA production to the high mutation rate the protein patterns were compared by two dimensional gel electrophoresis. Ten proteins were found to be differentially expressed at levels more than three fold greater in cells with than without msDNA, nine of which were identified by MALDI TOF MS. Eight of the nine identified proteins were repressed in msDNA-producing cells and, surprisingly, most were proteins functioning in the dissimilation of various carbon sources. One protein was induced four fold greater in the msDNA producing cells and was identified as a 30S ribosomal protein S2 involved in the regulation of translation. The molecular mechanism underlying the elevated mutation in msDNA-producing cell still remains elusive.

Tolerance of Anaerobic Granular Sludge to Oxygen

  • Shiru Jia;Benyi Xiao;Choi, Du-Bok;Cho, Ki-An;Kim, Young-Sik
    • 환경생물
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    • 제21권4호
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    • pp.405-409
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    • 2003
  • To study the tolerant capacity of anaerobic granular sludge (AGS) to oxygen using semi -dynamic batch experiment, the aerating time, pH of the basal media, reductive inorganic materials, microorganism, and microorganism metabolite were investigated. When the aerating time was higher or lower than 0.5 h, the producing gas activity of sludge was lower than that of the control. The oxygen tolerance of the experimental sludge was the highest at the initial pH 7.2. The producing gas activity of sludge I was higher than that of sludge II. And storage at $4^{\circ}C$ can low the lose of the oxygen tolerance capacity of granular sludge. The producing gas activity of sludge was the highest when KI was added. The growth of aerobic microorganisms and some metabolite could increase the producing gas activity of granular sludge.

Bacillus subtilis KL-57로부터 생산되는 생체계면활성제 합성 유전자 클로닝 (Cloning of Biosurfactant-Producing Gene from Bacillus subtilis KL-57)

  • 강상모;이병옥;이철수
    • 한국미생물·생명공학회지
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    • 제22권6호
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    • pp.593-598
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    • 1994
  • A bacterium KL-57 which exhibited biosurfactant activity was isolated. This bacterium was identified as Bacillus subtilis. The biosurfactant-producing gene of B. subtilis KL-57 was cloned into R subtilis MI113 by using plasmid pTB523. The plasmid DNA from the clone was found to carry a 18 kb PstI insert. The biosurfactant-producing gene was cleaved into 4 fragments by SmaI, 3 fragments by PvulI or EcoRl, 4 fragments by PvulI and EcoRI double digestion, 5 fragments by AccI, and 2 fragments by KpnI, HindIII or BamHI. By subcloning the 18 kb Pstl insert, a 2.3 kb EcoRl fragment conferred the biosurfactant producing activity on B. subtilis cells. The 2.3 kb had one HindIII cleave site. But Two fragments, which corresponds HindIII/EcoRl termini, exhibited no biosurfactant activity.

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버어리종 연초 곁순발생의 품종간 차이 (Varietal Difference of Sucker producing Characteristics in Burley Tobacco(Nicotiana tabacum L. var. burley))

  • 정석훈;조천준;최상주;조명조;백종운
    • 한국연초학회지
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    • 제22권1호
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    • pp.39-44
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    • 2000
  • This study was carried out to obtain the basic information about the sucker producing characteristics of certain burley tobacco varieties(Nicotiana tabacum L. var. burley) at Suwon and Chonju Experiment Station, Korea Ginseng & Tobacco Research Institute in 1999. The number and weight of suckers for 12 varieties or breeding lines were investigated three times, 45 days after transplanting for ground sucker, 10 days after topping, and 10 days after maleic hydrazide(MH) applying for upstalk sucker. KB 103 and KB 9416-1 produced less ground suckers and upstalk suckers than other entries did. KB 9210-8 produced less ground suckers and (Male sterile TC 612 x KB 108)F$_1$ also produced less upstalk suckers than other entries did. There was no differences among entries in sucker production at 10 days after MH treatment. Between the weight of total sucker and days to flower revealed the negative correlation, but not significant. The low sucker producing entries described above could be used as a parental line in the breeding program for the low sucker producing variety.

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Mycotoxin분비 수종균주들의 배양여과액으로 처리된 HeLa세포의 전자현미경적 관찰 (Electron Microscopec Observations on the HeLa Cells treated with culture Filtrates of Mycotoxin-Producing Fungi)

  • 고춘명;최대경;등영건
    • 미생물학회지
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    • 제12권3호
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    • pp.101-114
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    • 1974
  • The fine structure of HeLa cells treated with several mycotoxin-producing fungi (Aspergillus flavus ATCC 15517, Aspergillus parastiticus RIB 1037, Penicillium toxicarium RIB 4002, Penicillium cirinum SWU)238, Penicillium islandicum IFO 5235, Penicillium tadum IFO 5787 and Pencillium brunneum RIB 1172) has been examined and some details have been descried. The normal HeLa cell have numerous microvilli, large ovoid nucleus, pleomorphic mitochondria, electron-dense body, Golgi complex, mid-body and endoplasmic reticulum etc. Certain specific structural changes induced by culture filtrates of several mycotoxin-producing fungi have been noted. These alterations induced disappearance of Golgi complex, rER vacuolization, nucleolus attachment to the nuclear envelope nad appearance of certain vacuoles. There were not any changes by the treatment of culture filtrates of non-toxic fungi and only cell debris of some specimens can be observed by the injury of culture filtrates. The experimental animals treated with mycotoxin-producing fungi (Aspergillus flavus ATCC 15517, Aspergillus parasilicus RIB 1037, Penicillum citrinum SWU 238, Penicillium toxicarium RIB 4002, and Penicillium islandicum IFO 5235) were mal cells treated with culture filtrates.

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Use of Flp-Mediated Cassette Exchange in the Development of a CHO Cell Line Stably Producing Erythropoietin

  • Kim, Min-Soo;Lee, Gyun-Min
    • Journal of Microbiology and Biotechnology
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    • 제18권7호
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    • pp.1342-1351
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    • 2008
  • The feasibility of the use of Flp-mediated cassette exchange in the development of a CHO cell line, which produces erythropoietin (EPO) stably and largely, was investigated. A stable, high enhanced green fluorescence protein (EGFP)-producing clone was screened by extensive flow cytometric analysis. An EPO expression unit was targeted into the premarked locus of the stable parental clone by Flp-mediated cassette exchange and a correctly targeted clone (FC28T7) was obtained. The EPO production of FC28T7 was proven to be stable in long-term culture. Furthermore, the Flp-mediated cassette exchange did not alter the stable parental clone's characteristics concerning transgene expression level and stability. Taken together, the data obtained here indicated that the establishment of CHO cell lines stably producing a desired protein is achievable using Flp-mediated cassette exchange.

포유동물 세포와 생쥐 배에서 Retrovirus Vector를 이용한 SEAP와 GEP 유전자의 전이 및 발현 (Transfer and Expression of SEAP (secreted alkaline phosphatase) or GFP (green fluorescence protein) Gene in Mammalian Cells and Mouse Embryos by Using Retrovirus Vector System)

  • 김태완;이규승;박세필
    • 한국가축번식학회지
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    • 제20권3호
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    • pp.333-341
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    • 1996
  • One of the biggest problems involved in transgenic animal production is lack of appropriate market genes. To overcome this problem, we tested whether the genes of SEAP (secreted alkaline phosphatase) and GFP (green fluorescence protein) on our retrovirus vectors can be applicable to the transgenic animal production. The main advantage of these marker genes over other generally mainpulation can be selected without sacrificing viability. The results obtained in this study are summarized as follows: 1. Removal of zona pellucida from the mouse zygotes did not affect embryo developments to blastocysts. 2. Co-culture of zona-free embryos with virus-producing cells for 6 hours also did not affect embryo developments to blastocysts. 3. Among 58 blastocysts developed from the zona-free zygotes co-cultured with the virus-producing cells, SEAP expression was observed from the 6 blastocysts. 4. Expression of the GFP gene was detected from the virus- producing cells but no embryo expressing the gene was counted among 50 blastocysts developed from the zona-free zygotes co-cultured with the virus-producing cells.

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Molecular Characterization and Prevalence of 16S Ribosomal RNA Methylase Producing Bacteria in Amikacin Resistant Gram-negative Bacilli Isolated from Clinical Specimens

  • Shin, Kyung-A;Hwang, Seock-Yeon;Hong, Seung-Bok
    • 대한의생명과학회지
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    • 제18권3호
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    • pp.299-306
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    • 2012
  • Recently, the prevalence of 16S rRNA methylase conferring high-level resistance to aminoglycosides has been increasing in Gram-negative bacilli globally. We determined the prevalence and genotype of these methylase-producing bacteria, and characterized the co-resistance to ${\beta}$-lactam antibiotics and quinolone in Gram-negative clinical isolates collected in 2010 at a hospital in Korea. Among 65 amikacin-resistant isolates screened from 864 Gram-negative bacilli (GNB), 16S rRNA methylase genes were detected from 49 isolates, including Acinetobacter baumannii (43), Klebsiella pneumoniae (2), Proteus mirabilis (2) and Serratia marcescens (1), Empedobacter brevis (1). All of the 16S rRNA methylase genotype was armA and no variant sequences of amplified PCR products for armA were noted. The 16S rRNA methylase producing bacteria showed much higher resistance to aminoglycoside for Enterobacteriaceae and glucose non-fermenting (NF)-GNB and to imipenem for glucose NF-GNB, than the non-producing isolates. All of the 16S rRNA methylase producing Enterobacteriaceae had the extended-spectrum-${\beta}$-lactamase. In addition, two K. pneumoniae concurrently produced both plasmid-mediated AmpC ${\beta}$-lactamase and qnrB gene. All of the amikacin-resistant A. baumannii (43) co-harbored armA 16S rRNA methylase and $bla_{OXA-23}$ carbapenemase. In conclusion, 16S rRNA methylase producing bacteria were very prevalent among GNB in South Korea, and were commonly associated with co-resistance, including carbapenem and quinolone.

Evaluation of Various PCR Assays for Detection of Emetic-Toxin-Producing Bacillus cereus

  • Kim, Jung-Beom;Kim, Jae-Myung;Park, Yong-Bae;Han, Jeong-A;Lee, Soon-Ho;Kwak, Hyo-Sun;Hwang, In-Gyun;Yoon, Mi-Hye;Lee, Jong-Bok;Oh, Deog-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제20권7호
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    • pp.1107-1113
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    • 2010
  • Because conventional methods for detecting emetic-toxin-producing B. cereus are laborious and costly, various PCR assays, which are easy and cheap, have recently been reported. Therefore, this study estimated and compared the ability of various PCR assays to detect emetic-toxin-producing B. cereus strains isolated in Korea. The PCR assays were performed on 160 B. cereus strains, including 40 emetic-toxin-producing strains. Although the species-specific PCR assays were all shown to be highly specific, the sensitivities varied greatly. The accuracies of the primers were 97.5% (CER), 95.6% (EM1), 96.3% (RE234), 89.4% (CES), and 83.1% (Ces3R/CESR2). Moreover, the CER primer had a higher sensitivity (100%) than all the other primers tested, and a specificity of 96.7%. Thus, the CER primer was shown to be the most effective for screening the emetic-toxin-producing B. cereus strains tested in this study. However, the ability of these PCR assays to identify emetic-toxin-producing B. cereus should also be confirmed using other methods.