• Title/Summary/Keyword: Aspergillus nidulans

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Aspergillus nidulans 분비소낭 구성요소인 α-COP과 ε-COP의 결합 부위 분석 (Analysis of Protein Domain for Interaction between α-COP and ε-COP in Aspergillus nidulans)

  • 송은정;김기현;이환희;박정석;강은혜;박희문
    • 한국균학회지
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    • 제40권4호
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    • pp.224-228
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    • 2012
  • A. nidulans ${\alpha}$-COP과 상호작용하는 단백질을 동정하기 위하여 ${\alpha}$-COP을 암호하는 유전자를 bait로 yeast two-hybrid 스크리닝용 A. nidulans cDNA 라이브러리를 탐색한 결과, COPI 소낭의 구성요소 중 하나인 ${\varepsilon}$-COP을 암호화하고 있는 유전자를 동정하고 $aneA^+$($\underline{A}$spergillus $\underline{n}$idulans $\underline{e}$psilon-COP, $AN{\varepsilon}$-COP)으로 명명하였다. $aneA^+$ 유전자는 총 296개의 아미노산을 암호화하고 있으며, 다른 균류의 ${\varepsilon}$-COP과 높은 상동성을 보였다. Yeast two hybrid 시스템으로 두 단백질 간의 상호작용 부위를 분석한 결과, ${\alpha}$-COP의 COOH 도메인과 ${\varepsilon}$-COP의 C-말단부가 필수 부위였으며, ${\alpha}$-COP N-말단의 WD 도메인과 ${\varepsilon}$-COP의 TPR 부위는 두 단백질 간의 결합을 촉진하는 조절부위로 밝혀졌다. 또한 사상균인 A. nidulans와 효모류인 S. cerevisiae에서 ${\alpha}$-COP과 ${\varepsilon}$-COP 간 작용양상이 유사한 것으로 보아, COPI 소낭의 구성요소인 ${\alpha}$-COP과 ${\varepsilon}$-COP 간의 상호작용 기전은 진핵세포 내에서 진화적으로 잘 보존되어 있는 것으로 추정되었다.

Depletion of ${\varepsilon}$-COP in the COPI Vesicular Coat Reduces Cleistothecium Production in Aspergillus nidulans

  • Kang, Eun-Hye;Song, Eun-Jung;Kook, Jun Ho;Lee, Hwan-Hee;Jeong, Bo-Ri;Park, Hee-Moon
    • Mycobiology
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    • 제43권1호
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    • pp.31-36
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    • 2015
  • We have previously isolated ${\varepsilon}$-COP, the ${\alpha}$-COP interactor in COPI of Aspergillus nidulans, by yeast two-hybrid screening. To understand the function of ${\varepsilon}$-COP, the $aneA^+$ gene for ${\varepsilon}$-COP/AneA was deleted by homologous recombination using a gene-specific disruption cassette. Deletion of the ${\varepsilon}$-COP gene showed no detectable changes in vegetative growth or asexual development, but resulted in decrease in the production of the fruiting body, cleistothecium, under conditions favorable for sexual development. Unlike in the budding yeast Saccharomyces cerevisiae, in A. nidulans, over-expression of ${\varepsilon}$-COP did not rescue the thermo-sensitive growth defect of the ${\alpha}$-COP mutant at $42^{\circ}C$. Together, these data show that ${\varepsilon}$-COP is not essential for viability, but it plays a role in fruiting body formation in A. nidulans.

Aspergillus oryzae에서의 이종 Promoter들의 발현 (Expression of Heterologous Promoters in Aspersillus oryzae)

  • 함영태;김희정
    • KSBB Journal
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    • 제10권1호
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    • pp.38-45
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    • 1995
  • Aspergillus oryzae에서 A. nidulans의 glyceral d dehyde-3-phosphate dehydrogenase (gpdA)와 trpC, prmoter의 발현 능력 을 E. coli lacZ gene fusion을 이용하여 비교.분석하였다. A. oryzae 내에서 발현된 E. coli $\beta$galactosidase의 specific activIty를 조사하여 본 결과, gpdA promoter를 가지는 transformant들 에서는 2,000unit/ug of protem 정도의 activity를 보이는 반면, trpC, promater를 가지고 있는 transformant들에서는 10.5~52.3unit/ug of protein 정도의 activity를 보였다. 이 결과로부터 A. oryzae 내에서 A. nidulans의 gpdA promoter가 trpC, promoter에 비해 70 배 정도 더 강한 발현 능력을 보이고 있음을 알 수 있다. Western blot 분석에서도 gpdA promoter를 가지고 있는 transf ormant에서 더 많은 E. coli $\beta$-galactosidase가 발현된 것 을 보여 주고 있다. 또한 southern blot 분석에서는 이러한 강한 발현이 transform된 plasmid의 copy number와 상관 없음을 보여주고 있다.

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고농도 CO2 노출에 의한 Aspergillus nidulans의 유성생식 촉진효과 (Effect of High CO2 Concentration on Activation of Sexual Development in Aspergillus nidulans)

  • 한갑훈;양영석;김종화
    • 한국균학회지
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    • 제41권3호
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    • pp.192-196
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    • 2013
  • 진균의 분화과정은 다양한 환경요인에 의하여 영향을 받는다. 모델 사상성 진균인 Aspergillus nidulans의 경우 빛이 존재하거나 높은 염 농도, 비 발효성 당에 의하여 무성분화가 촉진되며 반대로 빛이 없거나 발효성 당이 풍부할 때, 그리고 저산소 조건일 경우 유성분화를 촉진하게 된다. 또한 veA나 nsdD와 같은 유성생식 양성조절유전자들도 유성분화를 유도하는 것으로 알려져 있다. 본 연구에서는 A. nidulans에서 $CO_2$의 농도와 분화패턴의 관계를 알아보고자 하였다. 정상적인 조건에서 $veA^+$ 야생형 균주는 유성생식과 무성생식이 균형을 이룬 상태의 분화를 진행하게 된다. 그러나 5% 정도의 높은 $CO_2$ 조건에서 배양하였을 경우 무성분화는 일어나지 않고 유성생식으로만 분화과정이 일어나게 된다. 뿐만 아니라 이러한 분화 양상은 veA와 nsdD에 의존적으로 일어나게 된다. 고농도의 $CO_2$ 조건이라 할지라도 $veA^-$ 혹은 $nsdD^-$ 돌연변이 균주에서는 유성분화는 일어나지 않고 무성분화만이 일어나는 것을 관찰할 수 있었는데, 이는 $CO_2$ 농도가 높아져도 유성생식에 있어서 이들 유전자의 기능이 필요하다는 것을 시사한다. 또한 5% $CO_2$ 조건은 인간 병원성 진균에게 있어서 사람의 신체 내에 살아남기 위하여 적응하여야 하는 대기 조건으로, 이러한 A. nidulans의 $CO_2$ 농도에 따른 분화양상의 변화는 A. fumigatus와 같은 인간 병원균의 생리, 분화적 변화에 대한 비교분석에 사용될 수 있다.

Aspergillus nidulans의한 경주마의 후당염 2예 (Two Cases of Guttural Pouch Mycosis in Race Horses Caused by Aspergillus nidulans)

  • 하태영;조길재;박응복;김상재
    • 한국임상수의학회지
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    • 제12권1호
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    • pp.853-860
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    • 1995
  • Two cases of guttural pouch mycosis in race horses were observed for clinical and pathological aspects of the disease to investigate etiology and pathogenesis of dysphagia and epistaxis of the horse. In case 1 showing prolonged dysphagic sign a diphtheritic membrane was confined to the guttural pouch involved with neuritis of the glossopharyngeal nerve due to fungal penetration. The other horse showing fatal recurrent epistaxis had lesion of mycetoma invading the internal carotid artery to provoke erosion of the artery. An Aspergillosis sp. was isolated from the guttural pouches of the two horses and identified as A nidulans.

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Promotion of Asexual Development and Inhibition of Sexual Development of Aspergillus nidulans by Short-Chain Primary Amines

  • Song, Myung-Hoon;Kuppusamy Selvam;Park, Chang-Jun;Jahng, Kwang-Yeop;Han, Dong-Min;Chae, Keon-Sang
    • Journal of Microbiology
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    • 제40권3호
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    • pp.230-233
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    • 2002
  • Effects of short-chain primary amines on Aspergillus nidulans development were analyzed. Propylamine induced asexual development and inhibited sexual development. Even on medium containing lactose as the sole carbon source, on which little conidial heads are formed and sexual structures are formed preferentially, or when sexual development was induced, propylamine induced asexual development and inhibited sexual development. These effects of propylamine seemed to be due to accumulation of mRNA of the brlA gene, which has been identified as a positive regulator of asexual development, and due to the reduction of the veA mRNA level. The veA gene has been identified as an activator of sexual development and also as an inhibitor of asexual development. Other primary amines, methylamine and ethylamine, showed identical effects on development where short-chain primary amino also promoted asexual development and inhibited sexual development.

Comparison of Cell Wall Ultrastructures of Aspergillus nidulans in Presence and Absence of a MnpAp Mannoprotein

  • Jeong, Hyo-Yong;Whang, Sung-Soo;Chae, Keon-Sang
    • Animal cells and systems
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    • 제10권3호
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    • pp.131-135
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    • 2006
  • The ultrastructure of Aspergillus nidulans cell wall in relation to a mannoprotein was studied by scanning and transmission electron microscopy. An mnpAp null-mutant, DMPV1, was used as a negative control of a wild type VER7. To analyze whether the mannoprotein in the cell wall during the development of an mnpAp null-mutant is present or not, immunogold microscopy was also adopted. The surface sculpturing of various cell types - hyphae, conidium, Hulle cell, and ascospore - were not very different between the wild type and the mnpAp-null mutant (DMPV1) as examined by scanning electron microscopy. These results were comparable to those examined by transmission electron microscopy, in that the hyphal cell wall was not indentical between two strains, probably caused by the MnpA protein (MnpAp). MnpAp was absent in both the hyphal cell wall of the DMPV1 strain and the conidial cell wall of a wide type, but clearly recognized in the hyphal cell wall of a wild type.

Microscopic Examination of the Suppressive Action of Antifungal Substances from Pseudomonas aeruginosa on Asexual Sporulation of Fungi

  • Kwon S. Yoon;Bu Y. Min;Park, Hyoung T.;Lee, Jong K.;Kim, Kun W.
    • Journal of Microbiology
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    • 제37권1호
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    • pp.27-34
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    • 1999
  • Two fractions with unusual antifungal activity that suppress asexual sporulation of several fungi were obtained from culture filtrate of Pseudomonas aeruginosa and were partially purified through the repeated silicagel flash column chromatographies. The sporulation-suppressive actions of these fractions in Aspergillus nidulans, Rhizopus stolonifer, and Coprinus cinereus, were analyzed by light and electron microscopes. The germination ability of the spores produced in the presence of these fractions were also checked to determine the persistent effects of these antifungal substances on the next generation. Light microscopic observation of developing sporangia of R. stolonifer grown in the presence of both fractions revealed that the significant number of sporangia failed to reach maturity, and frequently, uncontrolled growths of hyphae and rhizoids from the sporangiophores were found. In A. nidulans addition of these fractions appeared to cause different classes of morphological abnormality in conidia development, which included aborted formation of conidiogenous cells from the apex of conidiophores and enhanced hyphal growths either at the tip or middle of the conidiophores. Germination abilities of spores obtained from the cultures grown in the presence of antifungal fractions were 40∼60% in Aspergillus, 50∼80% in Coprinus (thallic spores), and 30∼40% in Rhizopus compared to those of normal spores.

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Aspergillus nidulans 온도감수성 돌연변이주의 분리 및 분석 (Isolation and Analysis of Temperature Sensitive Mutants in Aspergillus nidulans)

  • 박찬규;강현삼
    • 미생물학회지
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    • 제16권3호
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    • pp.111-121
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    • 1978
  • About 40 temperature-sensitive mutants have been isolated as a preliminary step to study the spore germination, the cell cycle, and the control of macromolecular synthesis in Aspergillus nidulans. To obtain temperature-sensitive mutants rapidly and effectively, the selective enrichment method using antifungal antibiotic nystatin was developed. Based on the data which had applied to the concentration of auxotrophic mutants by the earlier investigators, the optimal concentration and the time of treatment at the nonpermissive temeprature were determined as 50 to 100 units per ml and 4.5 hr., respectively. Out of 41 ts mutants assigned to the strain symbol PK, thirteen that seemed to be arrested at the earlystage of spore germination were subjected to the further cytological and genetic analysis. Elght of these mutants are able to form germ tube and five not. Staining with acid fuchsin for the 5PK strains shows that one irreversible mutant, PK6 strain able to form germ tube, accumulate mitotic spindle, being arrested in mitosis. Another PK15 and PK23 strain have more than one intact nucleolus without germ tube formation at the restrictive temperature. the temperature-senstive mutation in PK12 strain, the onlystrain which is able occurred in certain gene specific for the germination of spore. All of the ts markers are recessive and complement each other in heterokaryon between two different ts markers at the restrictive temperature.

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Aspergillus nidulans의 역제의 유전자의 작용기작에 관한 연구 II (Studies on the mechanisms of suppression in Aspergillus nidulans)

  • 문은표;강현삼
    • 미생물학회지
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    • 제20권1호
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    • pp.1-10
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    • 1982
  • From Aspergillus nidulans, six suppressor mutants, MSI-MS6, were isolated, and their characteristics were analysed. These were the suppressor mutants for acriflavin resistant and nicotinamide auxotrophic mutant phenotypes. MS1, MS2, MS5 and MS6, were linked to the chromosome IV, I, II respectively, and MS2 was linked to one of the rest chromosomes, MS3 and MS6 mutants had both suppressors for acriflavin resistant marker and for nicotinamide auxotrophic marker. In order to know the stability and efficiency of the suppressors, their reversion frequencies, that is, frequencies of losing the suppressibility, were analysed. Only MS3 and MS5 were reversed with high frequency. The four mutants didn't lose their suppressibilities, and this meant that the suppressors of these four were very stable and highly effcient. The suppressor specificities of these mutants were tested for other mutant's phenotype marker. One of the six suppressors, MS1, had the suppressor specificity for acriflavin resistant marker of 163 strain.

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