• Title/Summary/Keyword: URA3

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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.

Nucleotide Analysis of Phaffia rhodozyma DNA Fragment That Functions as ARS in Saccharomyces cerevisiae

  • Chung, Hee-Young;Hong, Min-Hee;Chun, Young-Hyun;Bai, Suk;Im, Suhn-Young;Lee, Hwanghee-Blaise;Park, Jong-Chun;Kim, Dong-Ho;Chun, Soon-Bai
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.650-655
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    • 1998
  • The chromosomal DNA fragment from Phaffia rhodozyma CBS 6938 which is able to autonomously replicate in the yeast Saccharomyces cerevisiae was cloned on an integrative URA3 plasmid. Its minimal fragment exhibiting autonomously replicating activiy in the S. cerevisiae gave a higher frequency transformation efficiency than that found for centromere-based plasmid, and enabled extrachromosoma1ly stable transmission of the plasmids in one copy per yeast cell under non-selective culture condition. The 836-bp DNA element lacked an ORF and did not contain any acceptable match to an ARS core consensus. Sequence analysis, however, displayed a cluster of three hairpin-Ioop-sequences with individual $\triangle {G_{25}}^{\circ}C$ free energy value of -10.0, -17.5, and -17.0 kcal. $mor^{-l}$as well as a 9-bp sequence with two base pair mismatches to the S. cerevisiae/E. coli gyrase-binding site. This 836-bp sequence also included one 7-bp sequence analogous to the core consensus of centromeric DNA element III (CDEIII) of S. cerevisiae, but CDEIII-like 7 bp sequence alone did not give a replicative function in this yeast.

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Effects of spThoc7 Deletion on Growth and mRNA Export in Fission Yeast (분열효모에서 spThoc7 유전자의 결실이 생장 및 mRNA Export에 미치는 영향)

  • Koh, Eun-Jin;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.249-253
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    • 2014
  • THOC7/Mft1 is a member of the THO complex that is an evolutionally conserved assembly connecting transcription elongation with mRNP packaging and mRNA export. In fission yeast Schizosaccharomyces pombe, an ortholog (spThoc7) of THOC7/Mft1 was isolated by partial complementation of the lethality in synthetic lethal mutant, SLRsm1. A deletion mutant in a diploid strain was constructed by replacing one of spThoc7-coding region with an $ura4^+$ gene using one-step gene disruption method. Tetrad analysis showed that the spthoc7 is nonessential for growth. But the ${\Delta}thoc7$ null mutant showed slight defects of both growth and mRNA export. And the functional spThoc7-GFP protein is localized mainly in the nucleus. These results suggest that spThoc7 is also involved in mRNA export from the nucleus to cytoplasm.

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.

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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Cotransformation of Saccharomyces cerevisiae with Yip and Yep Vectors (YEp 및 YIp 벡터에 의(依)한 Saccharomyces cerevisiae의 Cotransformation)

  • Lee, Sung Bum;Rhee, In Koo
    • Current Research on Agriculture and Life Sciences
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    • v.4
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    • pp.36-41
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    • 1986
  • The transformation of Saccharomyces cerevisiae with YIp26, YRp7 and YEp13 was investigated. Transformation frequences of YIp5, YIp26, YRp7 and YEp13 in Escherichia coli HB101 was $5.1{\times}10^{-4}$, $1.5 {\times}10^{-3}$, $1.3{\times}10^{-3}$, $3{\times}10^{-3}$, respectively. When plasmids were used in covalently closed circular form, transformation frequency of YEp13 was $1.2{\times}10^{-4}$ in S. cerevisiae DBY747 and $3.3{\times}10^{-4}$ in S. cerevisiae MC16 and that of YRp7, YIp26 was $3{\times}10^{-6}$, below $6{\times}10^{-8}$ respectively in S. cerevisiae DBY747 by the method of Ito. Cotransformation of YIp26 and YEp13 in linear form increased the frequency of transformation with efficiences 270-fold higher than transformation of YIp26 only in S. cerevisiae DBY747. In cotransformation of YIp5+YEp13 and YIp26+YRp7 with S. cerevisiae DBY747 by Beggs' method. Expression frequency of YIp5+YEp13 and YIp26+YRp7 was $4{\times}10^{-6}$, $1.5{\times}10^{-6}$, respectively. The recombinant plasmid of cotransformant was thought that YIp26 and YEp13, YIp5 and YEp13, and YIp26 and YRp 7 were ligated in vivo in S. cerevisiae DBY747.

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FURTHER PURIFIED GINSENG EXTRACT FRACTION (D-O-ANA) FOR INSULIN RELEASE AND ITS MODE OF ACTION COMPARED WITH THE ISOLATED RESIDUAL COMPONENTS (인삼성분 D-O-ANa이 인슐린 분비에 미치는 영향 및 작용기전에 관한연구)

  • KIMURA Masayasu;SUZUKI Jun;WAKI Isami;KIMURA Ikuko;TANAKA Osamu;MATSU-URA Hiromichi
    • Proceedings of the Ginseng society Conference
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    • 1984.09a
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    • pp.191-197
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    • 1984
  • A further purified fraction (D-O-ANa) was obtained from DPG 3-2 fraction of Ginseng Radix by complete removal of saponins, nucleosides, nucleic acid bases, amino acids, and sugars. D-O-ANa - induced insulin release was investigated to compare with that of DPG 3-2 and other isolated components. Among the sub fractions of DPG 3-2, D-O-ANa exhibited the most potent release of insulin with or without high concentrations of glucose, and it particularly enhanced the second phase of glucose-induced insulin release. DGP 3-2 potentiated significantly the glucose-induced insulin release from the isolated islets of diabetic mice at increasing concentrations of extracellular calcium ions (0.16 - 2.5 mM). A definite relationship was found between calcium $(^{45}Ca)$ uptake and insulin release. Ginsenoside $(G)-Rb_1\;and\;G-Rg_1$ did not enhance the glucose-induced insulin release. The effect of ginseng saponins was blocked by glucose (16.7 mM), being distinctly different from the glucose-potentiated effect of DPG 3-2. The insulin release effect of $G-Rg_1$ was unaffected by the presence or absence of extracellular $Ca^{2+}$ and theophylline. Adenosine also increased insulin release from isolated islets, but had no effect on perfused rat pancreas. Arginine stimulated insulin release less evidently than D-O-ANa, though arginineand adenosine-induced glucagon releases were more remarkable. In conclusion, D-O-ANa appears to be a major fraction in insulin release activity of ginseng and its mode of action may be related to $Ca^{2+}$ ion uptake. This physiological mechanism was distinct from that of the abnormal release induced by ginseng saponins.

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When Dose Losses of Maternal Lymphocytes Response to Trophoblast Antigen or Alloantigen Occur in Women with a History of Recurrent Spontaneous Abortion? (반복유산을 경험한 환자에서 임신중 태반항원과 동종항원에 노출된 모체 림프구면역반응은 언제부터 소실되나?)

  • Choi, Bum-Chae;Hill, Joseph A.
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.115-122
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    • 1998
  • The maintenance of a viable pregnancy has long been viewed as an immunological paradox. The deveolping embryo and trophoblast are immunologically foreign to the maternal immune system due to their maternally inherited genes products and tissue-specific differentiation antigens (Hill & Anderson, 1988). Therefore, speculation has arisen that spontaneous abortion may be caused by impaired maternal immune tolerance to the semiallogenic conceptus (Hill, 1990). Loss of recall antigen has been reported in immunosuppressed transplant recipients and is associated with graft survival (Muluk et al., 1991; Schulik et al., 1994). Progesterone $(10^{-5}M)$ has immunosuppressive capabilities (Szekeres-Bartho et al., 1985). Previous study showed that fertile women, but not women with unexplained recurrent abortion (URA), lose their immune response to recall antigens when pregnant (Bermas & Hill, 1997). Therefore, we hypothesized that immunosuppressive doses of progesterone may affect proliferative response of lymphocytes to trophoblast antigen and alloantigen. Proliferative responses using $^3H$-thymidine ($^3H$-TdR) incorporation of peripheral blood mononuclear cells (PBMCs) to the irradiated allogeneic periperal blood mononuclear cells as alloantigen, trophoblast extract and Flu as recall antigen, and PHA as mitogen were serially checked in 9 women who had experienced unexplained recurrent miscarriage. Progesterone vaginal suppositories (100mg b.i.d; Utrogestan, Organon) beginning 3 days after ovulation were given to 9 women with unexplained RSA who had prior evidence of Th1 immunity to trophoblast. We checked proliferation responses to conception cycle before and after progesterone supplementation once a week through the first 7 weeks of pregnancy. All patients of alloantigen and PHA had a positive proliferation response that occmed in the baseline phase. But 4 out of 9 patients (44.4%) of trophoblast antigen and Flu antigen had a positive proliferative response. The suppression of proliferation response to each antigen were started after proliferative phase and during pregnancy cycles. Our data demonstrated that since in vivo progesterone treated PBMCs suppressed more T-lymphocyte activation and $^3H$-TdR incorporation compare to PBMCs, which are not influenced by progesterone. This data suggested that it might be influenced by immunosuppressive effect of progesterone. In conclusion, progesterone may play an important immunological role in regulating local immune response in the fetal-placental unit. Furthermore, in the 9 women given progesterone during a conception cycle, Only two (22%) repeat pregnancy losses occured in these 9 women despite loss of antigen responsiveness (one chemical pregnancy loss and one loss at 8 weeks of growth which was karyotyped as a Trisomy 4). These finding suggested that pregnancy loss due to fetal aneuploidy is not associated with immunological phenomena.

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2, 4, 5-Trichlorophenoxyacetic Acid 분해균의 유전적 특성에 관한 연구

  • Yoon, So-Yeong;Son, Hong-Joo;Lee, Geon;Lee, Sang-Joon;Lee, Jong-Kun
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.260-264
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    • 1992
  • Pseudotnorju.c sp. EL-071P degrading 2.4.5-trichlorophe~~oxyi~cetaicci d (2.3.5-T) was resistantto antibiotics: rifampicin. ampicillin. kanamycin and metal ions : Zn" and Cu".The plasmitl related to the degradation of 2.4.5-'r and rifa~npicin resistance was isolatecifrom the strain. Its size was about 40 Kb. As result of transforming the plasmid intoEsch~rirhiti coli MClOhl, it was confirmed that the plasmid ura.; related to 2.4.5-T degradation.The strain coulil grow in the various chlorinated aromatic analogs as the solc carbon source.In the case of chlorophcnols. the chlorinated mono-substituteti phenols were easily dcgradetlin the order ol' ortho-. ~ ~ a r um- ,c ~tu-position.T he 2.3.5-T mctaholism was inhibited by 4-chlorophenol of 2.4.5-7' analog. In non-chlorinateci aromatics. ~ C I I L O ~ I ~ Csa.l icylilte i~ndtoluene were uscd ax the carbon source by the strain and typestrain Acudonlotrtr.\ plrtirltrKCTC 1643 having clegrad;~bility of various aromatics. But naphtalene was usecl only bythe A~urlomonri.\ sp. EL-07 1 P.the A~urlomonri.\ sp. EL-07 1 P.

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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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