• Title/Summary/Keyword: velA gene

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Effect of Various Carbon Sources on the Development of Aspergillus nidulans with $velA^+$ or velA1 allele (각종 탄소원이 $velA^+$ 및 velA1 Aspergillus nidulans의 분화에 미치는 영향)

  • Han, Dong-Min;Han, Yoo-Jeong;Chae, Keon-Sang;Jahng, Kwang-Yeop;Lee, Young-Hoon
    • The Korean Journal of Mycology
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    • v.22 no.4
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    • pp.332-337
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    • 1994
  • Under standard condition (Han, et al., 1990: glucose 1%-nitrate 0.1% minimal medium, 30 ml in 9 cm plate, $10^6$ cells of inoculum per plate), wild type of Aspergillus nidulans developed both sexual and asexual organs in ballance, while velA1 mutant developed asexual ones preferentially. Increase of glucose concentration did not significantly affect the asexual sporulation. However, development of sexual organs were largely affected. It was greatly enhanced when favorable nitrogen source, for example, casein hydrolysate was added, which is contrary to the case of Neurospora or Saccharomyces where limitation of N source induces sexual development. On most of moderate C sources asexual development in $velA^+$ strain was largely inhibited except acetate on which only asexual spores were produced, while that in velA1 mutant strain was not affected. Lactose promoted the sexual development even in velA1 mutant indicating that lactose itself or its metabolic intermediate may induce sexual development independent of allelic state of velA gene. On other moderate favorable C sources, glycerol, galactose and ethanol, asexual development was largely inhibited in $velA^+$ strain but not in velA1 mutant strain. Sexual organs were, however, never produced on acetate. These results suggested that asexual development of wild type is largely dependent on C sources and the velA gene is involved in the repression of asexual development in not-enough-grown (non-competent) thalli resulting in preferential progression of sexual development.

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Unveiling the Functions of the VosA-VelB Target Gene vidD in Aspergillus nidulans

  • Son, Ye-Eun;Park, Hee-Soo
    • Mycobiology
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    • v.49 no.3
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    • pp.258-266
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    • 2021
  • The velvet regulators VosA and VelB are primarily involved in spore maturation and dormancy. Previous studies found that the VosA-VelB hetero-complex coordinates certain target genes that are related to fungal differentiation and conidial maturation in Aspergillus nidulans. Here, we characterized the VosA/VelB-inhibited developmental gene vidD in A. nidulans. Phenotypic analyses demonstrated that the vidD deleted mutant exhibited defect fungal growth, a reduced number of conidia, and delayed formation of sexual fruiting bodies. The deletion of vidD decreased the amount of conidial trehalose, increased the sensitivity against heat stress, and reduced the conidial viability. Moreover, the absence of vidD resulted in increased production of sterigmatocystin. Together, these results show that VidD is required for proper fungal growth, development, and sterigmatocystin production in A. nidulans.

Construction of a High-Quality Yeast Two-Hybrid Library and Its Application in Identification of Interacting Proteins with Brn1 in Curvularia lunata

  • Gao, Jin-Xin;Jing, Jing;Yu, Chuan-Jin;Chen, Jie
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.108-114
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    • 2015
  • Curvularia lunata is an important maize foliar fungal pathogen that distributes widely in maize growing area in China, and several key pathogenic factors have been isolated. An yeast two-hybrid (Y2H) library is a very useful platform to further unravel novel pathogenic factors in C. lunata. To construct a high-quality full length-expression cDNA library from the C. lunata for application to pathogenesis-related protein-protein interaction screening, total RNA was extracted. The SMART (Switching Mechanism At 5' end of the RNA Transcript) technique was used for cDNA synthesis. Double-stranded cDNA was ligated into the pGADT7-Rec vector with Herring Testes Carrier DNA using homologous recombination method. The ligation mixture was transformed into competent yeast AH109 cells to construct the primary cDNA library. Eventually, a high qualitative library was successfully established according to an evaluation on quality. The transformation efficiency was about $6.39{\times}10^5$ transformants/$3{\mu}g$ pGADT7-Rec. The titer of the primary cDNA library was $2.5{\times}10^8cfu/mL$. The numbers for the cDNA library was $2.46{\times}10^5$. Randomly picked clones show that the recombination rate was 88.24%. Gel electrophoresis results indicated that the fragments ranged from 0.4 kb to 3.0 kb. Melanin synthesis protein Brn1 (1,3,8-hydroxynaphthalene reductase) was used as a "bait" to test the sufficiency of the Y2H library. As a result, a cDNA clone encoding VelB protein that was known to be involved in the regulation of diverse cellular processes, including control of secondary metabolism containing melanin and toxin production in many filamentous fungi was identified. Further study on the exact role of the VelB gene is underway.

Cloning of a Laccase Gene Fragment from Ganoderma lucidum (영지버섯 Laccase 유전자의 구리결합부위 I과 IV사이 지역의 클로닝)

  • 조지현;최형태;김경훈
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.192-195
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    • 2000
  • Degenerate primevs corresponding to the consensus sequences of the copper-binding regions in the N- and Cterminal domains of fungal laccases were used to isolate laccase gene-specific sequences froin a white rot rungus Ganodern~a lucidrm w-hich has been known to strengthen the imnnne system. A 1.6 Kbp fragment was amplified by PCR and its base sequence was detenuiued. Locating seven iutrous within the base sequence, we could deduce its amino acid sequence. The nucleotide sequence witl~out introlls was 47Y0 identical to that of lee1 gene of Pametes wllosa; lhe identity in amino acid sequences of the two was 7994 The deduced amino acid seqoence was also sunilar to those of Coriolus versicolo~ kc3 (79%); Co~,iolz~s hirsutus phenolouiduse (78%), Trainetes vel.srcoloi. lccl (77%), Trametes ~!i/Iosa Ice2 (77%) and Trametes vemicolor kc4 (66%).

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