• 제목/요약/키워드: URA3

검색결과 51건 처리시간 0.033초

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

  • 윤진호
    • 미생물학회지
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    • 제45권4호
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    • pp.300-305
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    • 2009
  • mRNA의 3' 말단형성 뿐만 아니라, 성숙한 mRNA의 핵에서 세포질로의 이동에 중요한 역할을 하는 출아효모 Saccharomyces cerevisiae의 폴리(A)-RNA 결합단백질인 Nab2와 유사한 분열효모 Schizosaccharomyces pombe의 단백질을 암호화하는 유전자(spNab2로 명명)의 결실돌연변이주(deletion mutant)를 제조하여 그 특성을 조사하였다. 이배체인 S. pombe 균주에 하나의 spNab2 유전자만을 결실시킨 후 4분체분석(tetrad analysis)을 수행한 결과, S. cerevisiae NAB2와는 다르게 이 유전자는 생장에 반드시 필요하지 않았다. 또한 spNab2 결실돌연변이는 mRNA의 핵에서 세포질로의 이동도 정상적으로 보였다. spNab2의 역할을 알아보기 위해, 티아민에 의해 발현이 조절되는 강력한 프러모터를 이용하여 spNab2를 과발현시켰다. spNab2 유전자가 과발현되면, 세포의 생장이 심하게 억제되었으며, 폴리(A)-RNA가 핵 안에 축적되고 세포질에서는 줄어들었다. 또한 GFP 융합단백질을 이용하여 spNab2 단백질의 세포 내 위치를 관찰한 결과, spNab2-GFP는 주로 핵 안에 존재하였지만 세포질에서도 관찰되었다. 이와 같은 결과들은 spNab2 유전자 역시 mRNA의 핵에서 세포질로의 이동에 관여하고 있음을 시사한다.

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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    • 제8권6호
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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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분열효모에서 spThoc7 유전자의 결실이 생장 및 mRNA Export에 미치는 영향 (Effects of spThoc7 Deletion on Growth and mRNA Export in Fission Yeast)

  • 고은진;윤진호
    • 미생물학회지
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    • 제50권3호
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    • pp.249-253
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    • 2014
  • THOC7/Mft1는 mRNA가 전사되는 동안 mRNP의 포장과 mRNA 방출에 관여하는 진화적으로 잘 보존된 THO 복합체의 구성인자이다. 분열효모 Schizosaccharomyces pombe에서 THOC7/Mft1의 이종상동체(spThoc7)가 합성치사 돌연변이체 SLRsm1의 생장 결함을 부분적으로 상보하는 것으로 선별되었다. 이배체 S. pombe 균주에 하나의 spthoc7 유전자만을 결실시킨 후 4분체 분석을 수행한 결과, 이 유전자는 생장에 필수적이지 않았다. 하지만, ${\Delta}thoc7$ 결실돌연변이는 생장과 mRNA의 핵에서 세포질로의 방출에 약간의 결함을 보였다. 기능을 하는 spThoc7-GFP단백질은 주로 핵 안에 존재하였다. 이와 같은 결과들은 spThoc7도 mRNA 방출에 관여하고 있음을 시사한다.

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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    • 제26권1호
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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
    • 한국추진공학회:학술대회논문집
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    • 한국추진공학회 2004년도 제22회 춘계학술대회논문집
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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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YEp 및 YIp 벡터에 의(依)한 Saccharomyces cerevisiae의 Cotransformation (Cotransformation of Saccharomyces cerevisiae with Yip and Yep Vectors)

  • 이승범;이인구
    • Current Research on Agriculture and Life Sciences
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    • 제4권
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    • pp.36-41
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    • 1986
  • 대장균(大腸菌)과 효모(酵母)의 셔틀 벡터인 YIp, YRp, YEp를 S. cerevisiae MC 16, DBY747에 형질전환(形質轉換)시켰다. 이들 벡터들을 대장균(大腸菌)에 형질전환(形質轉換)시켰을 때 그 빈도(頻度)가 YIp5, YIp26, YEp13, YRp7에서 각각 $5.1{\times}10^{-4}$, $1.5{\times}10^{-3}$, $3{\times}10^{-3}$, $1.3{\times}10^{-3}$으로 나타났다. YEp13을 MC16과 DBY747에 Ito 법(法)으로 형질전환(形質轉換)시켰을 때 MC16, DBY747에서의 빈도(頻度) 각각 $3.3{\times}10^{-4}$, $1.2{\times}10^{-4}$으로 나타났다. DBY747을 수용세포(受容細胞)로 하여 YRp7과 YIp26을 환상(環狀)으로 각각 형질전환(形質轉換)시켰을 때 그 빈도(頻度)는 각각 $3{\times}10^{-6}$, $6{\times}10^{-8}$이하로 나타났다. DBY747을 수용세포(受容細胞)로 하여 YEp13과 YIp26을 선상(線狀)으로 절단하여 $Li^+$ 처리(處理)한 S. cerevisiae에 cotransformation을 하였을 때 YIp26+YEp13 ($Leu^+$, $Ura^+$)의 cotransformant는 $1{\times}10^{-5}$ 빈도(頻度)가 나왔으며 YIp26과 YEp13에 대한 각각의 빈도(頻度)는 $10^{-7}$ 이하, $5{\times}10^{-5}$ 빈도(頻度)로 나타났다. 또 YIp5와 YEp13, YIp26과 YRp7을 원형질체화(原形質體化)한 S. cerevisiae DBY747에 cotransformation 하였을 때 빈도(頻度)는 YIp5+YEp13은 $4{\times}10^{-6}$으로 나오고 YIp26+YRp7에서 $1.5{\times}10^{-6}$으로 나타났다.

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

  • KIMURA Masayasu;SUZUKI Jun;WAKI Isami;KIMURA Ikuko;TANAKA Osamu;MATSU-URA Hiromichi
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1984년도 학술대회지
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    • pp.191-197
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    • 1984
  • 인삼의 저혈당성 분획 (DPG-3-2)이 적출한 랑겔한스섬에서 인슐린 분비를 증가시킨다고 일전에 본 저자들이 발표하였다. 근래에 이와 같은 DPG-3-2 분획으로부터 사포닌, 뉴크레오사이드, 뉴크레오타이드, 아미노산, 당 등을 완전히 제거시켜 더욱 정제한 D-O-ANa 성분을 추출하였다. D-O-ANa 유발성 인슐린 분비효과를 DPG 3-2 분획과 그외 잔여성분과 비교 검토하였다. D-O-ANa는 글루코우즈 농도의 고저에 무관하게 인슐린 분비를 가장 강하게 촉진하였다. 특히 D-O-ANa는 제 2 차증 글루코운즈 유발성 인슐린 분비를 촉진하였다. DPG 3-2 분획은 당뇨병 쥐로부터 떼어낸 랑겔한스섬에서 세포외 체액의 칼슘이온이 증가됨에 따라($0.16{\~}6.25$mM) 글루코우즈 유발성 인슐린 분비를 더욱 현저히 증가시켰다. 칼슘이온 흡수와 인슐린분비의 상호관계가 뚜렷이 밝혀졌다. 이 관계는 single sucrose gap 방법에 의해 주의 적출 간 문맥에서 칼슘주파수가 증가되는 실험을 통해 증명되었다.

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

  • 최범채
    • Clinical and Experimental Reproductive Medicine
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    • 제25권2호
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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

  • 윤소영;송홍주;이건;이상준;이종근
    • 미생물학회지
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    • 제30권4호
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    • pp.260-264
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    • 1992
  • 2, 4, 5-T 분해균주인 Pseudomonas sp. EL-071P 는 rifampicin, ampicilin, kanamycin 등의 중금속에 대하여 내성을 가지고 있었다. 이 균주로부터 2, 4, 5-T 분해능과 rifampicin 의 내성에 관련된 하나의 plasmid 를 분리, 정제하였으며 그 크기는 약 40 kb 이었다. E. coli 에 2, 4, 5-T 분해능에 관ㅎ련된 plasmid 를 transformation 한 결과 2, 4, 5-T 분해에는 plasmid 가 관여함을 확인할 수 있었다. 이 균주는 다양한 염소계방향족 동족체화합물을 유일한 탄소원으로 하여 생육할 수 있었으며, chlorophenol compound 의 경우 CI$^{-}$ 의 치환위치가 ortho, para-, meta- 위치순으로 분해가 잘 되었다. 2, 4, 5, -T 의 기질 유사체인 4-chlorophenol 에 의해 2, 4, 5-T 대사가 저해되었다. 비염소계 방향족화합물중 benzoate, salicylate, toluene 은 Pseudomonas sp. EL-071P 와 Pseudomonas putida KCTC 1643 에 의해 탄소원으로 이용되었으나 naphtalene 의 경우 Pseudomonas sp. EL-071 에 의해서만 탄소원으로 이용되어 표준균주인 Pseudomonas putida KCTC 1643 과 상이한 결과를 나타내었다.

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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- (THE OPEN-ORIENTED CHARACTERISTICS OF THE NEW TOWN HOUSE 'MACHIYA' IN KYOTO FROM ITS 'HUKI-NUKE' SPACE POINT OF VIEW)

  • 김태영
    • 건축역사연구
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    • 제5권1호
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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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