• 제목/요약/키워드: complementation

검색결과 299건 처리시간 0.039초

Methylovorus sp. Strain SS1 DSM11726으로부터 rpoH 유전자의 클로닝과 염기서열 분석 (Cloning and Nucleotide Sequence Analysis of the rpoH Gene from Methylovorus sp. Strain SS1 DSM11726)

  • 엄치용;송승은;박미화;김영민
    • 한국미생물·생명공학회지
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    • 제35권3호
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    • pp.177-183
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    • 2007
  • 열충격 시그마인자를 코딩하는 유전자 rpoH가 결여된 돌연변이체 대장균(Escherichia coli satrain A7448)을, 메탄올 자화세균인 Methylovorus sp. strain SS1 DSM11726의 phagemid library로 형질전환 시켜서 $30^{\circ}C$에서 성장하는 Escherichia coli strain A7448 로부터 Methylovorus sp. strain SS1 DSM11726의 rpoH 유전자를 클로닝하고 그 염기서열을 분석하였다. 1,793-bp 염기서열 분석 결과 Methylovorus sp. strain SS1 DSM11726의 RpoH는 284개의 아미노산으로 이루어져 있었으며 예상된 분자량은 32,006, p1값은 5.79로 나타났으며, 동일계열의 ${\beta}$-proteobacteria에 속하는 세균들의 RpoH와 높은 상동성을 보여주었다. Methylovorus sp. strain SS1 DSM11726의 RpoH는 대장균의 RpoH의 기능을 대신할 수 있음을 보여주었다. 열충격 후 RpoH양은 15분까지 지속적으로 증가하다 20분 뒤 양이 감소하는 양상을 나타내었다. 이는 Methylovorus sp. strain SS1 DSM11726의 RpoH 단백질 역시 열에 의해 유도됨을 말해 준다.

Cell Death Mediated by Vibrio parahaemolyticus Type III Secretion System 1 Is Dependent on ERK1/2 MAPK, but Independent of Caspases

  • Yang, Yu-Jin;Lee, Na-Kyung;Lee, Na-Yeon;Lee, Jong-Woong;Park, Soon-Jung
    • Journal of Microbiology and Biotechnology
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    • 제21권9호
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    • pp.903-913
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    • 2011
  • Vibrio parahaemolyticus, which causes gastroenteritis, wound infection, and septicemia, has two sets of type III secretion systems (TTSS), TTSS1 and TTSS2. A TTSS1-deficient vcrD1 mutant of V. parahaemolyticus showed an attenuated cytotoxicity against HEp-2 cells, and a significant reduction in mouse lethality, which were both restored by complementation with the intact vcrD1 gene. V. parahaemolyticus also triggered phosphorylation of mitogen-activated protein kinases (MAPKs) including p38 and ERK1/2 in HEp-2 cells. The ability to activate p38 and ERK1/2 was significantly affected in a TTSS1-deficient vcrD1 mutant. Experiments using MAPK inhibitors showed that p38 and ERK1/2 MAPKs are involved in V. parahaemolyticus-induced death of HEp-2 cells. In addition, caspase-3 and caspase-9 were processed into active forms in V. parahaemolyticus-exposed HEp-2 cells, but activation of caspases was not essential for V. parahaemolyticus-induced death of HEp-2 cells, as shown by both annexin V staining and lactate dehydrogenase release assays. We conclude that secreted protein(s) of TTSS1 play an important role in activation of p38 and ERK1/2 in HEp-2 cells that eventually leads to cell death via a caspase-independent mechanism.

Molecular Characterization of Adenylyl Cyclase Complex Proteins Using Versatile Protein-Tagging Plasmid Systems in Cryptococcus neoformans

  • So, Yee-Seul;Yang, Dong-Hoon;Jung, Kwang-Woo;Huh, Won-Ki;Bahn, Yong-Sun
    • Journal of Microbiology and Biotechnology
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    • 제27권2호
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    • pp.357-364
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    • 2017
  • In this study, we aimed to generate a series of versatile tagging plasmids that can be used in diverse molecular biological studies of the fungal pathogen Cryptococcus neoformans. We constructed 12 plasmids that can be used to tag a protein of interest with a GFP, mCherry, $4{\times}FLAG$, or $6{\times}HA$, along with nourseothricin-, neomycin-, or hygromycin-resistant selection markers. Using this tagging plasmid set, we explored the adenylyl cyclase complex (ACC), consisting of adenylyl cyclase (Cac1) and its associated protein Aca1, in the cAMP-signaling pathway, which is critical for the pathogenicity of C. neoformans. We found that Cac1-mCherry and Aca1-GFP were mainly colocalized as punctate forms in the cell membrane and non-nuclear cellular organelles. We also demonstrated that Cac1 and Aca1 interacted in vivo by co-immunoprecipitation, using $Cac1-6{\times}HA$ and $Aca1-4{\times}FLAG$ tagging strains. Bimolecular fluorescence complementation further confirmed the in vivo interaction of Cac1 and Aca1 in live cells. Finally, protein pull-down experiments using $aca1{\Delta}$::ACA1-GFP and $aca1{\Delta}$::ACA1-GFP $cac1{\Delta}$ strains and comparative mass spectrometry analysis identified Cac1 and a number of other novel ACC-interacting proteins. Thus, this versatile tagging plasmid system will facilitate diverse mechanistic studies in C. neoformans and further our understanding of its biology.

Oxidative Damage of DNA Induced by Ferritin and Hydrogen Peroxide

  • Kang, Jung-Hoon
    • Bulletin of the Korean Chemical Society
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    • 제31권10호
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    • pp.2873-2876
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    • 2010
  • Excess free iron generates oxidative stress that may contribute to the pathogenesis of various causes of neurodegenerative diseases. Previous studies have shown that one of the primary causes of increased brain iron may be the release of excess iron from intracellular iron storage molecules. In this study, we attempted to characterize the oxidative damage of DNA induced by the reaction of ferritin with $H_2O_2$. When DNA was incubated with ferritin and $H_2O_2$, DNA strand breakage increased in a time-dependent manner. Hydroxyl radical scavengers strongly inhibited the ferritin/$H_2O_2$ system-induced DNA cleavage. We investigated the generation of hydroxyl radical in the reaction of ferritin with $H_2O_2$ using a chromogen, 2,2'-azinobis-(2-ethylbenzthiazoline-6-sulfonate) (ABTS), which reacted with ${\cdot}OH$ to form $ABTS^{+\cdot}$. The initial rate of $ABTS^{+\cdot}$ formation increased as a function of incubation time. These results suggest that DNA strand breakage is mediated in the reaction of ferritin with $H_2O_2$ via the generation of hydroxyl radicals. The iron-specific chelator, deferoxamine, also inhibited DNA cleavage. Spectrophotometric study using a color reagent showed that the release of iron from $H_2O_2$-treated ferritin increased in a time-dependent manner. Ferritin enhanced mutation of the lacZ' gene in the presence of $H_2O_2$ when measured as a loss of $\alpha$-complementation. These results indicate that ferritin/$H_2O_2$ system-mediated DNA cleavage and mutation may be attributable to hydroxyl radical generation via a Fenton-like reaction of free iron ions released from oxidatively damaged ferritin.

주거공간에 적용 가능한 신소재 특성 및 기준에 관한 기초 연구 (A Preliminary Study on the Characteristics and Criteria of New Material for the Possibility of Application in Housing Space)

  • 정선희;서지은
    • 한국주거학회논문집
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    • 제21권4호
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    • pp.71-79
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    • 2010
  • This study is a fundamental research which suggests some standards that can be used in selecting some advanced materials to be applied in housing space designs. The following conclusions have been drawn from the study. First, it has been discovered that as the residents' quality of life improved, the finishing materials emerged as an important element of housing spaces. It could also be seen that among the finishing materials used in housing spaces, there is recently a growing trend towards various environmentally-friendly advanced materials under the theme of 'the nature' and 'health'. As a result, the need to investigate the standards of discriminatory and efficient housing space designs using advanced materials is on the increase. Second, through the literature and precedent studies, the physical criteria of advanced materials have been classified into five categories: new technology, new manufacturing methods, new materials, complementation and applicability. The main criteria of advanced finishing materials for housing spaces have been classified into four main categories: durability, functionality, economical efficiency and sensibility. By collecting and classifying some key words according to each of these four categories, the representative or combinable features have been extracted. By organizing the advanced materials' physical criteria and their details, some standards based on the physical aspects have been suggested. The features of the finishing materials in consideration of advanced materials have been reclassified into the user-based criteria, environmental criteria and spatial criteria. Through this method, the criteria have been brought up from a new viewpoint, establishing the standards for the advanced materials applicable to housing spaces. Finally, after assessing the criteria by analyzing the advanced materials currently known to be applicable in housing spaces, it has been concluded that the criteria can be used as the fundamental standards. As this study is based on investigating the literature and data, it may be less than reasonable to generalize from the limited number of examples. Hence, it shall be important to make a profound study of some detailed criteria by suggesting more objective standards and analyzing a greater variety of advanced materials.

코르셋 룩 원명에 관한 연구 - 20대 전기 표준체형 여성을 중심으로 - (A Study on Corset Look Pattern - Focused on Women in Their 20s with Standard Body Type -)

  • 김시만;성옥진
    • 복식
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    • 제57권8호
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    • pp.48-59
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    • 2007
  • The purpose of this research is to develop a basic pattern of corset look taking into consideration characteristics of human body and fit, by transforming 3 dimensional information obtained from draping of standard body type of women in their 20s, which constitute the biggest consumer group of corset look in Korea, on the basis of body shape of women best reflecting 3 dimensional characteristics. In order to develop a corset look pattern suitable for standard body type of women in their 20s, wear tests of A, B, and C types of patterns in terms of the bust in the standard body type that bust circumference 86.7cm and chest circumference 87.0cm were evaluated best. Taking into consideration above findings, fer the purpose of developing a corset look pattern having an excellent fit and reflecting characteristics of Korean women in their 20s with standard body type, a corset look pattern was designed with proper ease calculated by draping incorporated, and through alteration and complementation of such a pattern, a study pattern was produced. This study is expected to provide tips for standard body type women in their 20s with smaller bust circumference to choose clothing allowing for a better fit able to make up for weakness of their body, while presenting to clothing makers ways of designing and size-setting in terms of characteristics of body type of these consumers.

Deletion of GBG1/AYR1 Alters Cell Wall Biogenesis in Saccharomyces cerevisiae

  • Ahn, Ki-Woong;Kim, Sung-Woo;Kang, Hyung-Gyoo;Kim, Ki-Hyun;Park, Yun-Hee;Choi, Won-Ja;Park, Hee-Moon
    • Mycobiology
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    • 제38권2호
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    • pp.102-107
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    • 2010
  • We identified a gene for $\beta$-1,3-glucan synthesis (GBG1), a nonessential gene whose disruption alters cell wall synthesis enzyme activities and cell wall composition. This gene was cloned by functional complementation of defects in $\beta$-1,3-glucan synthase activity of the the previously isolated Saccharomyces cerevisiae mutant LP0353, which displays a number of cell wall defects at restrictive temperature. Disruption of the GBG1 gene did not affect cell viability or growth rate, but did cause alterations in cell wall synthesis enzyme activities: reduction of $\beta$-1,3-glucan synthase and chitin synthase III activities as well as increased chitin synthase I and II activities. GBG1 disruption also showed altered cell wall composition as well as susceptibility toward cell wall inhibitors such as Zymolyase, Calcofluor white, and Nikkomycin Z. These results indicate that GBG1 plays a role in cell wall biogenesis in S. cerevisiae.

Neurospora crassa 유전자에 의한 Saccharomyces cerevisiae coq7 돌연변이의 회복 (Restoration of Saccharomyces cerevisiae coq7 Mutant by a Neurospora crassa Gene)

  • 김은정;김상래;이병욱
    • 생명과학회지
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    • 제13권6호
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    • pp.933-942
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    • 2003
  • Coenzyme Q은 긴 isoprenoid 사슬을 갖는 quinone의 유도체이다. Coenzyme Q는 진핵생명체의 미토콘드리아의 내막과 원핵생명체의 세포막에 위치하는 전자전달계에 존재하는 지용성 물질이며, 또한 항산화제로의 기능도 갖는다. Coenzyme Q는 Saccharomyces cerevisiae의 호기적 성장에 필수적이며, coq 돌연변이체는 발효가 불가능한 탄소 원에서의 성장이 불가능하다. S. cerevisiae의 $coq^7$p 효소들과 유사성을 나타내는 단백질을 암호화하는 Neurcspora crassa cDNA를 효모의 발현 벡터에 삽입하였다. N. crassa COQ7의 예상 서열은 S. cerevisiae의 효소와 58% homology를 보였다. N. crassa $coq^{-7}$ 유전자의 S. cerevisiae $coq^7$ 형질전환체는 야생형 균주와 유사한 성장률을 보였다. 형질전환 균주들은 발효가 불가능한 탄소원인 글리세롤을 유일한 탄소원으로 배양하였을 경우에도 정상적인 성장을 나타냈다. 또한 불포화지방산인 linolenic acid를 성장 배지에 첨가하여도 야생형 균주와 유사한 생존율이 관찰되었다.

치마버섯에서 이형 유전자 발현에 관한 연구 (The Studies on the Heterogenous Gene Expression in Schizophyllum commune)

  • 박동철;김현정;김옥미;배준태;박선희;이병훈;이갑랑
    • 한국균학회지
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    • 제26권1호통권84호
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    • pp.103-107
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    • 1998
  • Coprinus cinereus에서 tryptophan 생합성에 관여하는 trp2 유전자의 이종 버섯에서의 발현여부를 조사하였다. 유전자 상동성을 확인하기 위하여, C. cinereus trp2 유전자를 함유하는 재조합 DNA인 pHIONA8을 probe로 사용하여 southern blot을 한 결과 S. commune 1-71의 tryptophan 생합성 유전자와 상당한 homology가 있는 것으로 나타났다. pHIONA8 DNA를 이용한 S. commune T1(tryptophan 영양요구주)의 상보성 검증에서는 pertri-dish당 약 50개의 형질전환체가 나타났으며, 이들 형질전환체는 다른 S. commune의 다른 화합성 균주와의 교배 실험에서 clamp cell의 유도와 동시에 약 $2{\sim}3$주만에 자실체를 관찰하였다.

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과시소비성향집단별 패션관여가 자기만족도에 미치는 영향 (The Effects of Fashion Involvement on Self-Satisfaction by Conspicuous Consumption Tendency Groups)

  • 박현주;박숙현
    • 한국의류학회지
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    • 제36권11호
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    • pp.1162-1173
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    • 2012
  • This study classifies conspicuous consumption groups and the difference of fashion involvement and selfsatisfaction by each group. It also examined the effect of fashion involvement on self-satisfaction by each group. A questionnaire method was used for the study method and the subjects of the study were females in their 20s-50s. A total of 580 sets of questionnaires were distributed and 554 sets were used for the final analysis. Data were analyzed by factor analysis, t-test, ANOVA, factor analysis, cluster analysis, Cronbach's alpha coefficients, and multiple regression analysis. The results of this study were as follows: First, this study classified 4 groups of active conspicuous consumption, the group of passive conspicuous consumption, the group of syntonic conspicuous consumption and the group pursuing individuality & frugal consumption. Second, as a result of the examination of the impact of fashion involvement for each group with a propensity for conspicuous consumption on their self-satisfaction, it was found that the sex appeal of fashion involvement had no significant impact on the economic satisfaction in the group of active conspicuous consumption, and had no significant impact on all elements of self-satisfaction in the group of passive conspicuous consumption. It was also found that social symbolism had a negative impact on satisfaction with looks in the group of syntonic conspicuous consumption, and the physical complementation and directions of looks had a negative impact on satisfaction with living, the social symbolism on satisfaction with looks and the syntone on satisfaction with looks in the group of pursuing individuality & frugal consumption.