• Title, Summary, Keyword: Aspergillus nidulans

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Expression of Aspergillus awamori Glucoamylase Gene in Asperillus nidulans (Aspergillus nidulans내에서 Aspergillus awamori의 Glucoamylase 유전자 발현)

  • 김석준;유준희;정구홍
    • Korean Journal of Microbiology
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    • v.31 no.2
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    • pp.136-140
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    • 1993
  • The A. nidulans expression vector which contained trpC marker gene from A. nidulans was constructed to produce glucoamy]ase. The recombinant plasmid was introduced into auxotrophic mutant A. nidulans B17. Southern blot analysis of the genomic DNA from transformant showed that pKHG2 DNA had integrated into the A. nidulans chromosomes. Northern analysis of the total RNA from transform ant showed that mRNA of glucoamylase gene was synthesized in induction condition. Specific activity of glucoamylase was increased in transform ants. G]ucoamylase was shown to be active in non-denaturing acrylamide gel.

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Studies on the Organization and Transcription of Aspergillus nidulans tRNA Genes (Aspergillus nidulans의 tRNA 유전자의 구성과 발현에 관한 연구 II. Aspergillus nidulans 총 tRNA 유전자의 cloning)

  • 이병재;강현삼
    • Korean Journal of Microbiology
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    • v.21 no.4
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    • pp.229-237
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    • 1983
  • Total tRNA genes from Aspergillus nidulans were cloned for the further investigation of the structure and expression of Aspergillus tRNA genes. Aspergillus DNA was isolated from spores and cloned into pBR322 plasmid DNA using BamHI and $T_4$ ligase. The recombinant hybrid DNA was transformed into E. coli HB101 and some 30,000 transformants were initially selected. Of these, about 5,300 E. coli clones containing Aspergillus DNA inserted into plasmid pBR322 at BamHl site have been isolated. The hybridization data obtained from the labeled Aspergillus $^{32}P-tRNA$ indicated that 105 colonies carried the total tRNA genes. From the data above and cohybridization experiment, tRNA genes of Aspergillus nidulans seem to be twice more clustered than those of yeast.

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Conserved Roles of MonA in Fungal Growth and Development in Aspergillus Species

  • Son, Ye-Eun;Park, Hee-Soo
    • Mycobiology
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    • v.47 no.4
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    • pp.457-465
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    • 2019
  • MonA is a subunit of a guanine nucleotide exchange factor that is important for vacuole passing and autophagy processes in eukaryotes. In this study, we characterized the function of MonA, an orthologue of Saccharomyces cerevisiae Mon1, in the model fungus Aspergillus nidulans and a toxigenic fungus A. flavus. In A. nidulans, the absence of AnimonA led to decreased fungal growth, reduced asexual reproduction, and defective cleistothecia production. In addition, AnimonA deletion mutants exhibited decreased spore viability, had reduced trehalose contents in conidia, and were sensitive to thermal stress. In A. flavus, deletion of AflmonA caused decreased fungal growth and defective production of asexual spores and sclerotia structures. Moreover, the absence of monA affected vacuole morphology in both species. Taken together, these results indicate that MonA plays conserved roles in controlling fungal growth, development and vacuole morphology in A. nidulans and A. flavus.

UVSC of Aspergillus nidulans is a Functional Homolog of RAD51 in Yeast

  • Yoon, Jin-Ho;Seong, Kye-Yong;Chae, Suhn-Kee;Kang, Hyen-Sam
    • BMB Reports
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    • v.34 no.5
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    • pp.428-433
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    • 2001
  • A defect in uvsC of Aspergillus nidulans caused high methyl methansulfonate (MMS)-sensitivity, hyporecombination, and a lack of UV induced mutation. The uvsC gene of Aspergillus nidulans shares a sequence similarity with the RAD51 gene of Saccharomyces cerevisiae. In this study, in vitro and in vivo tests were conducted in order to determine whether or not the UVSC protein had functional similarities to RAD51, the recombination enzyme in yeast. The purified recombinant UVSC protein, following expression in Escherichia coli, showed binding activity to single-stranded DNA (ssDNA), when both ATP and magnesium are present. In addition, ATPase activity was also demonstrated and its activity was stimulated in the presence of ssDNA. The UVSC protein that was expressed under the ADH promoter in S. cerevisiae suppressed in part the sensitivity to MMS of the rad51 null mutant. Similarly, when the uvsC cDNA was expressed from the nmt promoter, the MMS sensitivity of the rhp51 null mutant of Schizosaccharomyces pombe was partially complemented. These results indicate that the A. nidulans UVSC protein is a functional homologue of the RAD51 protein.

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Controlled Expression and Secretion of Aspergillus oryzae Alkaline Protease in Aspergillus nidulans

  • Kim, Eun-Ah;Lee, Jeong-Goo;Whang, Mi-Kyung;Park, Hee-Moon;Kim, Jeong-Yoon;Chae, Suhn-Kee;Maeng, Pil-Jae
    • Journal of Microbiology
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    • v.39 no.2
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    • pp.95-101
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    • 2001
  • In an effort to develop an efficient expression and secretion system for heterologous proteins in Aspergilius nidulans, the PCR-amplified coding sequence for alkaline pretense (AlpA) of A. oryzae was cloned into a fungal expression vector downstream of A. nidulans aicA (alcohol dehydrogenase) promoter to yield pRAAlp. Transformation of A. nidulans with pRAAlp gave stable transformants harboring various copy numbers (3 to 10) of integrated alpA gene, from among which 6 representatives were selected. On a medium containing 0.8% ammonium sulfate that represses the expression of the host's own pretense, the alcA prumoter-controlled AlpA expression was strongly induced by threonine but repressed by glucose. The level of AlpA secretion was highest (approximately 666 mU/ml) in transformant ALP6 containing the largest copy number integrated alpA. However, the level of AlpA secretion was not necessarily proportional to the copy numbers of the integrated alpA genes. The N-terminal sequence or the secreted mature AlpA was determined to be Gly-Leu-Thr-Thr-Gln-Lys-Ser and its molecular mass to be approximately 34 kDa, indicating that AlpA is properly processed by the removal of 121 N-terminal amino acids.

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Role of Sugars in Early Stage of Spore Germination in Filamentous Fungi, Aspergillus nidulans (사상균인 Aspergillus nidulans의 무성포자 발아와 당의 역할)

  • Chung, Kwang-Hee;Kim, Jae Won
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.511-518
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    • 2018
  • Initiation of spore germination in filamentous fungi such as Aspergillus nidulans and Botrytis cinerea requires the presence of nutrients. In this study, involvement of sugar sensing machinery was suggested in the germination of A. nidulans spores. Germination did not occur when the spores of A. nidulans were incubated in distilled water, whereas they were successfully germinated in the presence of 5% glucose with a germination rate of over 98% after 6hr incubation. Similar results were obtained when the spores were incubated in the presence of various sugars such as fructose, sucrose, and starch. Interestingly, spore germination was not observed in the presence of D-arabinose, whereas L-arabinose could induce germination as determined by the formation of germ tubes, indicating the presence of sugar sensing machinery that distinguish between the enantiomers of sugars. This inference was further supported by a decrease in germination rate (less than 25%) upon treatment of spores with trypsin. Subsequent MALDI-TOF mass spectrometry analysis of the surface proteins of spores identified ten proteins among which eight were involved in sugar metabolism. Taken together, our results suggest that spore germination in A. nidulans is initiated by the interaction of sugars with sugar binding proteins on the surface of spores.

Synergistic Effect of Substrates on the Biosynthesis of Cellulase and Xylanase Complexes from Aspergillus nidulans (Aspergillus nidulans 의 섬유질 분해효소계 생합성에 미치는 기질의 공조효과)

  • Lee, Jeong-Ae;Maeng, Jin-Soo;Maeng, Pil-Jae;Rhee, Young-Ha
    • The Korean Journal of Mycology
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    • v.17 no.2
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    • pp.57-65
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    • 1989
  • The effect of various cellulosic and hemicellulosic substrates on the induction of cellulase and xylanase complexes in Aapergillus nidulans was investigated. The most efficient substrates for the induction of cellulase and xylanase complexes were carboxymethylcellulose for endoglucanase, cellobiose for ${\beta}-glucosidase$, and xylan for endoxylanase and ${\beta}-xylosidase$, respectively. However, the mixtures of these substrates, especially CMC-xylan and CMC-xylan-laminarin mixture, were much more effective not only for the enhancement of the biosynthesis of all the cellulase and xylanase complexes but also for the balanced production of these enzyme components than individual substrate. The polyacrylamide gel electrophoresis followed by activity staining showed the variation in the patterns and relative intensity of ${\beta}-glucosidase$, endoglucanase and endoxylanase components in individual enzyme preparations from A. nidulans cultures grown on different substrates. These results suggest that the biosynthesis is of cellulase and xylanase systems in A. nidulans is regulated in coordination at the level of induction.

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A Novel Rapid Fungal Promoter Analysis System Using the Phosphopantetheinyl Transferase Gene, npgA, in Aspergillus nidulans

  • Song, Ha-Yeon;Choi, Dahye;Han, Dong-Min;Kim, Dae-Hyuk;Kim, Jung-Mi
    • Mycobiology
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    • v.46 no.4
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    • pp.429-439
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    • 2018
  • To develop a convenient promoter analysis system for fungi, a null-pigment mutant (NPG) of Aspergillus nidulans was used with the 4'-phosphopantetheinyl transferase (PPTase) gene, npgA, which restores the normal pigmentation in A. nidulans, as a new reporter gene. The functional organization of serially deleted promoter regions of the A. nidulans trpC gene and the Cryphonectria parasitica crp gene in filamentous fungi was representatively investigated to establish a novel fungal promoter assay system that depends on color complementation of the NPG mutant with the PPTase npgA gene. Several promoter regions of the trpC and crp genes were fused to the npgA gene containing the 1,034-bp open reading frame and the 966-bp 3' downstream region from the TAA, and the constructed fusions were introduced into the NPG mutant in A. nidulans to evaluate color recovery due to the transcriptional activity of the sequence elements. Serial deletion of the trpC and crp promoter regions in this PPTase reporter assay system reaffirmed results in previous reports by using the fungal transformation step without a laborious verification process. This approach suggests a more rapid and convenient system than conventional analyses for fungal gene expression studies.

Cell Cycle-dependent Expression of Chitin Synthase Genes in Aspergillus nidulans

  • Park, Bum-Chan;Maeng, Pil-Jae;Park, Hee-Moon
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.74-78
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    • 2001
  • The transcription of the chitin synthase genes (chss) was cell cycle-regulated in Aspergillus nidulans and the expression pattern was classified into two groups. Group one, containing chsA and chsC, showed decreasing transcription level upon entry into the S-phase and no further variation during the remainder of the cell cycle. However, group two, containing chsB, chsD, and csmA showed a sharp decrease of mRNA level upon entry into the G2-phase and an increase during the M-phase. Our results suggested that the chss, belonging to same group with the similar expression pattern during the cell cycle are functionally linked and that chsD may play a role in hyphal growth and development in A. nidulans.

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Ultrastructural Study on the Cleistothecium Development in Aspergillus nidulans

  • Sohn, K.T.;Yoon, K.S.
    • Mycobiology
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    • v.30 no.3
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    • pp.117-127
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    • 2002
  • Cleistothecial development in Aspergillus nidulans(teleomorph, Emericella nidulans) was examined with the transmission electron microscopy. Cleistothecial initial was a small coiled lump of cells, ca. 6 ${\mu}m$ in diameter, which was consisted of a slightly swollen core with a short "tail" hypha. Initials were wrapped with a loose layer of hyphae. Core cells of cleistothecial initials were broad and multinucleated at first, then formed dikaryotic ascogenous cells, followed by post-meiotic tetra-nucleate or octa-nucleate protoasci and finally mature ascospores. Croziers were formed early during cleistothecium development. The peridial layer of mature cleistothecia was derived from the wrapping hyphae which originally invested the young cleistothecium. Completion of peridial layers development was associated with the depositing of a non-enzyme reactive material around peridial cells. $H\ddot{u}lle$ cell formation during the cleistothecial development appeared to be somewhat coordinated with the developmental stages of cleistothecium.