• Title/Summary/Keyword: URA

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Characteristics of Antifungal Bacterium, Bacillus subtilis YS1 and It′s Mutant Induced by Gamma Radiation (온천수로부터 분리한 항진균세균의 특성 및 감마선$(Co^{60})$ 조사를 이용한 돌연변이체 유기)

  • 이영근;김재성;송인근;정혜영;장화형
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.305-311
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    • 2001
  • Antifungal bacterium, Bacillus subtilis YS1 was isolated from Yusong hot spring showed broad antifungal spectrum against 12 kinds of plant pathogenic fungi and Candida albicans, animal pathogen. From the gamma($Co^{60}$) radiation sensitivity test, $D_10$ value was 2.08 kGy and it survived above 20 kGy of radiation dose. Several mutants were induced by gamma radiation. Among them, YS1-1009 mutant showed resistance against tebuconazole of herbicide, increased activity against Botryoshaeria dothidea and ligninase activity. YS67 mutant was antifungal deficient auxotrophic mutants(trp-pro-or arg-ura-). From this results, it suggested that gamma irradiation could be useful method for mutant induction.

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Development of a Coded-aperture Gamma Camera for Monitoring of Radioactive Materials (방사성 물질 감시를 위한 부호화 구경 감마카메라 개발)

  • Cho, Gye-Seong;Shin, Hyung-Joo;Chi, Yong-Ki;Yoon, Jeong-Hyoun
    • Journal of Radiation Protection and Research
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    • v.29 no.4
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    • pp.257-261
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    • 2004
  • A coded-aperture gamma camera was developed to increase the sensitivity of a pin hole camera made with a pixellated CsI(Tl) scintillator and a position-sensitive photomultiplier tube. The modified round-hole uniformly redundant array of pixel size $13{\times}11$ was chosen as a coded mask considering the detector spatial resolution. The performance of the coded-aperture camera was compared with the pin hole camera using various forms of Tc-99m source to see the improvement of signal-to-noise ratio or the improvement of the sensitivity. The image quality is much improved despite of a slight degradation of the spatial resolution. Though the camera and the test were made for low energy case, but the concept of the coded-aperture gamma camera could be effectively used for the radioactive environmental monitoring and other applications.

Fission Yeast-based Screening to Identify Putative HDAC Inhibitors Using a Telomeric Reporter Strain

  • Chung, Kyung-Sook;Ahn, Jiwon;Choi, Chung-Hae;Yim, Nam Hui;Kang, Chang-Mo;Kim, Chun-Ho;Lee, Kyeong;Park, Hee-Moon;Song, Kyung-Bin;Won, Misun
    • Molecules and Cells
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    • v.26 no.1
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    • pp.93-99
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    • 2008
  • Transcriptional silencing is regulated by promoter methylation and histone modifications such as methylation and acetylation. We constructed a Schizosaccaromyces pombe reporter strain, KCT120a, to identify modifiers of transcriptional silencing, by inserting the $ura4^+$ gene into a heterochromatic telomere region. Two compounds inhibited the activity of histone deacetylases, induced acetylation of histone H3 and caused apoptotic cell death in HeLa cells. Expression of gelsolin and $p21^{waf1/cip1}$ also increased, as it does in response to HDAC inhibitors such as TSA. Therefore, these compounds appear to be potent inhibitors of HDACs, and hence potential anti-cancer drugs. Our observations suggest that a yeast cell-based assay system for transcriptional silencing may be useful for identifying histone deacetylase inhibitors and other agents affecting chromatin remodeling.

Development of a Genome-Wide Random Mutagenesis System Using Proofreading-Deficient DNA Polymerase ${\delta}$ in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Oh Cheol;Kim, Sang-Yoon;Hwang, Dong Hyeon;Oh, Doo-Byoung;Kang, Hyun Ah;Kwon, Ohsuk
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.304-312
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    • 2013
  • The thermotolerant methylotrophic yeast Hansenula polymorpha is attracting interest as a potential strain for the production of recombinant proteins and biofuels. However, only limited numbers of genome engineering tools are currently available for H. polymorpha. In the present study, we identified the HpPOL3 gene encoding the catalytic subunit of DNA polymerase ${\delta}$ of H. polymorpha and mutated the sequence encoding conserved amino acid residues that are important for its proofreading 3'${\rightarrow}$5' exonuclease activity. The resulting $HpPOL3^*$ gene encoding the error-prone proofreading-deficient DNA polymerase ${\delta}$ was cloned under a methanol oxidase promoter to construct the mutator plasmid pHIF8, which also contains additional elements for site-specific chromosomal integration, selection, and excision. In a H. polymorpha mutator strain chromosomally integrated with pHIF8, a $URA3^-$ mutant resistant to 5-fluoroorotic acid was generated at a 50-fold higher frequency than in the wild-type strain, due to the dominant negative expression of $HpPOL3^*$. Moreover, after obtaining the desired mutant, the mutator allele was readily removed from the chromosome by homologous recombination to avoid the uncontrolled accumulation of additional mutations. Our mutator system, which depends on the accumulation of random mutations that are incorporated during DNA replication, will be useful to generate strains with mutant phenotypes, especially those related to unknown or multiple genes on the chromosome.

Thrust Characteristics of a Laser-Assisted Pulsed Plasma Thruster

  • Masatoshi Kawakami;Hideyuki Horisawa;Kim, Itsuro ura
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 2004.03a
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    • pp.294-299
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    • 2004
  • An assessment of a novel laser-electric hybrid propulsion system was conducted, in which a laser-induced plasma was induced through laser beam irradiation onto a solid target and accelerated by electrical means instead of direct acceleration only by using a laser beam. A fundamental study of newly developed rectangular laser-assisted pulsed-plasma thruster (PPT) was conducted. On discharge characteristics and thrust performances with increased peak current compared to our previous study to increase effects of electromagnetic forces on plasma acceleration. Maximum peak current increased for our early study by increasing electromagnetic effects in a laser assisted PPT. At 8.65 J discharge energy, the maximum current reached about 8000 A. Plasma behaviors emitted from a thruster in various cases were observed with an ICCD camera. It was shown that the plasma behaviors were almost identical between low and high voltage cases in initial several hundred nanoseconds, however, plasma emission with longer duration was observed in higher voltage cases. Canted current sheet structures were also observed in the higher voltage cases using a larger capacitor. With a newly developed torsion-balance type thrust stand, thrust performances of laser assisted PPT could be estimated. The impulse bit and specific impulse linearly increased. On the other hand, coupling coefficient and the thrust efficiency did not increase linearly. The coupling coefficient decreased with energy showing maximum value (20.8 ?Nsec/J) at 0 J, or in a pure laser ablation cases. Thrust efficiency first decreased with energy from 0 to 1.4 J and then increased linearly with energy from 1.4 J to 8.6 J. At 8.65 J operation, impulse bit of 38.1 ?Nsec, specific impulse of 3791 sec, thrust efficiency of 8 %, and coupling coefficient of 4.3 ?Nsec/J were obtained.

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Effects of spNab2 Deletion and Over-Expression on mRNA Export (분열효모에서 spNab2 유전자의 결실돌연변이 및 과발현에 대한 분석)

  • Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.300-305
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    • 2009
  • We constructed the deletion mutants of fission yeast Schizosaccharomyces pombe spNab2 gene that is homologous to poly(A)-binding protein NAB2 in budding yeast Saccharomyces cerevisiae, which plays crucial roles in mRNA 3' end formation and mRNA export from nucleus into the cytoplasm. A null mutant in an $h^+$/ $h^+$ diploid strain was constructed by replacing the spNab2-coding region with an $ura4^+$ gene using one-step gene disruption method. Tetrad analysis showed that the spNab2 is not essential for vegetative growth and mRNA export. However, over-expression of spNab2 cause the severe growth defects and intensive accumulation of poly(A) RNA in the nucleus. Also, the spNab2-GFP fusions were localized mainly in the nucleus. These results suggest that spNab2 is also involved in mRNA export out of the nucleus.

Creation of an Ethanol-Tolerant Yeast Strain by Genome Reconstruction Based on Chromosome Splitting Technology

  • Park, A-Hwang;Sugiyama, Minetaka;Harashima, Satoshi;Kim, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.184-189
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    • 2012
  • We sought to breed an industrially useful yeast strain, specifically an ethanol-tolerant yeast strain that would be optimal for ethanol production, using a novel breeding method, called genome reconstruction, based on chromosome splitting technology. To induce genome reconstruction, Saccharomyces cerevisiae strain SH6310, which contains 31 chromosomes including 12 artificial mini-chromosomes, was continuously cultivated in YPD medium containing 6% to 10% ethanol for 33 days. The 12 mini-chromosomes can be randomly or specifically lost because they do not contain any genes that are essential under high-level ethanol conditions. The strains selected by inducing genome reconstruction grew about ten times more than SH6310 in 8% ethanol. To determine the effect of mini-chromosome loss on the ethanol tolerance phenotype, PCR and Southern hybridization were performed to detect the remaining mini-chromosomes. These analyses revealed the loss of mini-chromosomes no. 11 and no. 12. Mini-chromosome no. 11 contains ten genes (YKL225W, PAU16, YKL223W, YKL222C, MCH2, FRE2, COS9, SRY1, JEN1, URA1) and no. 12 contains fifteen genes (YHL050C, YKL050W-A, YHL049C, YHL048C-A, COS8, YHLComega1, ARN2, YHL046W-A, PAU13, YHL045W, YHL044W, ECM34, YHL042W, YHL041W, ARN1). We assumed that the loss of these genes resulted in the ethanol-tolerant phenotype and expect that this genome reconstruction method will be a feasible new alternative for strain improvement.

A Study on Dye Elution from the Circle Contact Lenses (써클콘택트렌즈의 염료 용출에 대한 연구)

  • Kim, So Ra;Kang, Ura;Seo, Bo Min;Park, Mijung
    • Journal of Korean Ophthalmic Optics Society
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    • v.19 no.2
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    • pp.171-177
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    • 2014
  • Purpose: The present study was conducted to investigate whether the elution of the dyes from the circle contact lenses existed or not when rubbed with cotton swab. Methods: The change of lens surface was observed by scanning electron microscope when a total of seven kinds of circle contact lens were rubbed with cotton swab, and the dye elution was further compared when the lenses were soaked in multi-purpose solution in $20^{\circ}C$ and $36.5^{\circ}C$ for a week. Results: Five of a total of 7 circle contact lenses showed the dye elution from the front or the back surface of the lens when rubbed with cotton swabs, and existence of dye elution was varied depending on the type of contact lenses and different at the front and back surface even in the same lens. The tear-off of the pigmented portion by the rubbing was found through scanning electron microscopic observation when the dye elution existed. However, the dye was not eluted when the circle contact lenses was soaked in a multi-purpose solution in $20^{\circ}C$ or $36.5^{\circ}C$ for a week. Ingredient of colorant was not provided by the manufacturers in more than 50% of the circle contact lenses investigated for this study and the basic information such as the pigmentation method was provided inappropriately. Conclusions: In this study, the result was the dye elution from circle contact lenses was obtained when a physical force is applied thus, it may appear to contribute the development of circle contact lenses and safety of lens wear.

THE OPEN-ORIENTED CHARACTERISTICS OF THE NEW TOWN HOUSE 'MACHIYA' IN KYOTO FROM ITS 'HUKI-NUKE' SPACE POINT OF VIEW (일본국(日本國) 경도(京都)의 정가(町家) 유형(類型) 연구(硏究) -신경정가(新京町家)의 "후끼누께" (취발(吹拔))를 통하여 본 "열림" 성향(性向)에 대하여-)

  • Kim, Tai-Young
    • Journal of architectural history
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    • v.5 no.1 s.9
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    • pp.50-72
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    • 1996
  • The typical Japanese house has the characteristics of open dwellings to make them cool in the hot and humid summer. And then the traditional town house 'Machiya', being built very closely to each other and walled up both sides, it has taken the open-oriented characteristics in itself. The purpose of this study is to clarify that the open-oriented of traditional 'Machiya' has been succeeded to the new 'Machiya' in the latest. The new 'Machiya' shows the open-oriented, taking the ventilative 'Huki-nuke' space of traditional 'Machiya' as the new spatial formal elements, in the changes such as scale material space organization. The characteristics of 'Huki-nuke' space are represented as follows ; 1. The facade of the traditional 'Machiya', which has taken on a semitransparent qualty, has been generally changed to the closing qualty except for the open parts of shop and garage. This facade of the new 'Machiya' has been taken to be in keeping with the existing town as much as possible. 2. A series of three rooms, composed of shop/living dining kitchen/room from the road, have been dispersed to every floors in a building with a very extensive scale. But this serial and linear type remains as the loosefit space, and the long dwellings of the upper stories are divided by type each dwelling unit. 3. 'Tori-niwa', which is a consecutive and penetrating space, connects the road with the rooms of dwelling and functions as the circulation of man thing energy, The new 'Machiya' changed to the multi-story, the corridor and the stair have been fumed up as the elements in the place of 'Tori-niwa' The 'Huki-nuke' space was locted in the hall, stairwell, living dining kitchen room, and so on. 4. The small court yard 'Tsubo-niwa' and back yard 'Ura-niwa' at the both ends of living spaces would be made a hole in a series of rooms and enclosed by the neighboring 'Machiya'. On the contrary the new 'Machiya' at present takes in the private and closing organization enclosing the innercourt. 5. The open-oriented ${\ulcorner}$In${\lrcorner}$ or ${\ulcorner}$Out${\lrcorner}$ is not brought out because of the delicate spatial formal configuration in the traditional 'Machiya'. But the open-oriented ${\ulcorner}$In${\lrcorner}$, all sides being closed by walls, is well brought out in the new 'Machiya'.

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Characterization of ${\alpha}$-amylase Producing Hybrid Constructed between Saccharomycopsis and Saccharomyces (Saccharomycopsis속과 Saccharomyces속의 잡종형성 균주에서 생산하는 ${\alpha}$-amylase의 특성)

  • Yang, Young-Ki;Moon, Myeng-Nim;Lim, Chae-Young;Rhee, Young-Ha;Kim, Jeong-Ho
    • Korean Journal of Microbiology
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    • v.35 no.4
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    • pp.315-321
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    • 1999
  • This study has been performed to deveope a yeast strain having high ${\alpha}$-amylase production ability using nuclear transfer method. Hybrids formed between the strains of Saccharomyces fiburigera KCTC 7393 and Saccharomyces cerevisiae KCTC 7049 (tyr-, ura-)were obtained by nuclear transfer technique. Nuclei isolated from the wild type S. fiburigera strain were transfered into auxotrophic mutants S. cerevisiae and selected the hybrids showing an increased starch degrading capability were selected (MN-16). This transformant grew best and produced maximal ${\alpha}$-amylase activity on the medium containing 2% (V/V) soluble starch. ${\alpha}$-Amylase from MN-16 was purified electrophoretically homogenety and its properties were investigated. The enzyme was purified about 10.6 fold with an overall yield 9.7% from the culture medium by ammonium sulfate fractionation. DEAE-Sephacel column chromatography, and Sephacryl S-200 column chromatography. The purified enzyme showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight of the ${\alpha}$-amylase was estimated to be 53,000 daltons by SDS-PAGE and by gel permeation chromatography on Sephacryl S-200. The purified enzyme showed the maximum activity at pH 5.5 and 40${\circ}C$. The km value for soluble starch was 2.5㎎/㎖. The enzyme activity increased in the presence of $Ca^{2+}, Co^{2+}, EDTA, Mg^{2+}, Mn^{2+}, Zn^{2+}$, but inhibited by $Cu^{2+}, Fe^{2+}$, and $Ni^{2+}$

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