• 한국양식학회:학술대회논문집
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• 한국양식학회 2006년도 수산관련학회 공동학술대회 발표요지집
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• pp.476-478
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• 2006

To analyze subcellular localization of betanodavirus protein B2, a plasmid expressing Betanodavirus protein B2 fused to enhanced green fluorescent protein (EGFP-Nl) was constructed. The transient expression of full-length B2 fused to EGFP in GF cells confirmed the equal distribution of protein B2 between cytoplasm and nucleus. However, transfection of N-terminal half of the B2 revealed that this truncated form predominantly localized to the cytoplasm. By using several deletion mutants and point mutants, we determined the regions and/or motif responsible for the subcellular localization of betanodavirus.

• Journal of Microbiology and Biotechnology
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• 제8권4호
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• pp.422-425
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• 1998

Chitin synthase null-mutants propagate in yeast form in RPMI medium with suppression of hyphal growth. This hyphal suppression is also observed in the wild type culture grown in RPMI medium supplemented with deer antler extract. To identify the possible target of deer antler extract, the enzymatic activities of chitin synthases were examined. The enzymatic activities of three chitin synthases, CAChsl, CAChs2, and CAChs3, were found to be differentially inhibited by deer antler extract. Of them, CAChsl, was the most sensitive to the extract. These results indicate that deer antler extract causes hyphal suppression, which resembles the effects of chitin synthase deletion, probably through direct inhibition of chitin synthases.

• Mycobiology
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• 제47권4호
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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.

• BMB Reports
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• 제41권11호
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• pp.778-783
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• 2008

Turnip yellow mosaic virus (TYMV) RNA has two hairpins in its 5' untranslated region (5'-UTR). To investigate the role of the hairpins in replication of TYMV, mutants lacking one or both of the two hairpins were constructed. The TYMV constructs were introduced into Chinese cabbage by an Agrobacterium-mediated T-DNA transfer method, called agroinfiltration. Analysis of total RNA from agroinfiltrated leaves showed that replication of the mutant TYMV RNA lacking both hairpins was about 1/100 of wild type. This mutant was also impaired in systemic spread. Deletion analysis of each hairpin revealed that both hairpins were needed for maximal replication. The deletion analysis along with sequence modification of the hairpin structure indicates that the second hairpin plays a role in efficient long-distance systemic movement of TYMV.

• 미생물학회지
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• 제42권2호
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• pp.153-155
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• 2006

mRNA의 핵에서 세포질로의 이동에 관여하는 발아효모 Saccharomyces cerevisiae의 SAC3유전자와 유사한 분열효모 Schizosaccharomyces pombe의 유전자 (spSac3로 명명)의 결실돌연변이주 (knockout mutant)를 제조하여 그 특성을 조사하였다. S. pombe의 이배체 (diploid) 균주에서 한 spSac3 유전자만을 결실시킨 후 4분체분석 (tetrad analysis)을 수행한 결과, 이 유전자는 성장에 필수적이었다. 또한 in situ hybridization을 통해 세포 내의 poly(A)+ RNA분포를 살펴본 결과, spSac3 유전자의 결실돌연변이주는 mRNA의 핵에서 세포질로의 이동에 이상이 있었다. 이와 같은 결과들은 spSac3 유전자 역시 mRNA의 핵에서 세포질로의 이동에 매우 중요한 역할을 담당하고 있음을 시사한다.

• ALGAE
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• 제24권2호
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• pp.85-91
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• 2009

The filamentous cyanobacterium Anabaena sp. Strain PCC 7120 produces a developmental patten of single hete- rocysts separated by approximately 10 vegetative cells. Heterocysts differentiate from vegetative cells and are spe- cialized for nitrogen fixation. The patS gene, which encodes a small peptide that inhibits heterocyst differentiation, is expressed in proheterocysts and plays a critical role in establishing the heterocyst pattem. Another key regulator of heterocyst development is the hetR gene. hetR mutants fail to produce heterocysts and extra copies of hetR on a plas- mid cause a multiple contiguous heterocyst phenotype. To elucidate the relationship between these two counter act- ing factors in the genetic regulatory pathway during heterocyst differentiation, the expression patterns of a patS-gfp and a hetR-gfp fusion were examined in a patS deletion and a hetR deletion strain. The results, in combination with the result from a hetR and patS double deletion strain, suggest patS and hetR are mutually antagonistic and the bal- ance between these two factors in tow different cell types (heterocysts and vegetative cells) may be critical during the decision making process on their cell fates.

• Mycobiology
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• 제41권3호
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• pp.145-148
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• 2013

Vegetative growth signaling of the opportunistic human pathogenic fungus Aspergillus fumigatus is mediated by GpaA ($G{\alpha}$). FlbA is a regulator of G protein signaling, which attenuates GpaA-mediated growth signaling in this fungus. The flbA deletion (${\Delta}flbA$) and the constitutively active GpaA ($GpaA^{Q204L}$) mutants exhibit enhanced proliferation, precocious autolysis, and reduced asexual sporulation. In this study, we demonstrate that both mutants also show enhanced tolerance against $H_2O_2$ and their radial growth was approximately 1.6 fold higher than that of wild type (WT) in medium with 10 mM $H_2O_2$. We performed quantitative PCR (qRT-PCR) for examination of mRNA levels of three catalase encoding genes (catA, cat1, and cat2) in WT and the two mutants. According to the results, while levels of spore-specific catA mRNA were comparable among the three strains, cat1 and cat2 mRNA levels were significantly higher in the two mutants than in WT. In particular, the ${\Delta}flbA$ mutant showed significantly enhanced and prolonged expression of cat1 and precocious expression of cat2. In accordance with this result, activity of the Cat1 protein in the ${\Delta}flbA$ mutant was higher than that of $gpaA^{Q204L}$ and WT strains. For activity of the Cat2 protein, both mutants began to show enhanced activity at 48 and 72 hr of growth compared to WT. These results lead to the conclusion that GpaA activates expression and activity of cat1 and cat2, whereas FlbA plays an antagonistic role in control of catalases, leading to balanced responses to neutralizing the toxicity of reactive oxygen species.

• The Plant Pathology Journal
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• 제22권3호
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• pp.215-221
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• 2006

Gibberella zeae (anamorph: Fusarium graminearum) is an important pathogen of cereal crops. This fungus produces a broad range of secondary metabolites, including polyketides such as aurofusarin (a red pigment) and zearalenone (an estrogenic mycotoxin), which are important mycological characteristics of this species. A screen of G. zeae insertional mutants, generated using a restriction enzyme-mediated integration (REMI) procedure, led to the isolation of a mutant (Z43R606) that produced neither aurofusarin nor zearalenone yet showed normal female fertility and virulence on host plants. Outcrossing analysis confirmed that both the albino and zearalenone-deficient mutations are linked to the insertional vector in Z43R606. Molecular characterization of Z43R606 revealed a deletion of at least 220 kb of the genome at the vector insertion site, including the gene clusters required for the biosynthesis of aurofusarin and zearalenone, respectively. A re-creation of the insertional event of Z43R606 in the wild-type strain demonstrated that the 220-kb deletion is responsible for the phenotypic changes in Z43R606 and that a large region of genomic DNA can be efficiently deleted in G. zeae by double homologous recombination. The results showed that 52 putative genes located in the deleted genomic region are not essential for phenotypes other than the production of both aurofusarin and zearalenone. This is the first report of the molecular characterization of a large genomic deletion in G. zeae mediated by the REMI procedure.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2005년도 제20차 학술대회 논문집
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• pp.34-34
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• 2005

• International Journal of Industrial Entomology and Biomaterials
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• 제6권1호
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• pp.81-85
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• 2003

To verify importance of the intervening sequence between the ribosome binding site (RBS) and the initiation codon for expression of Bacillus thuringiensis $\delta$-endotoxin, the pProMu, containing SphI and NcoIsites between RBS and the initiation codon of the cry1Ac gene, and the deletion derivatives of pProMu were constructed and transformed into the B. thuringiensis subsp. kurstaki $Cry^{-B}$ strain. The pProMu-ΔSphIhad identical six bases of intervening sequence to pProAc though the arrangement of sequence was different. Other mutants containing pProMu had 1 or 10 or 14 bases between RBS and the initiation codon. Among deletion mutants, only ProMu-ΔSphI/CB only produced 130 kDa typical bipyramidal crystals like those seen for ProAc/CB. However, ProMu/CB, $ProMu-{\Delta}NcoI$, and ProMu-ΔSphI＋NcoIdid not produce Cry1Ac crystals. In conclusion, the results suggest that 6-base intervening sequence was important for expression of cry1-type class gene. Furthermore, spacing effect of the intervening sequences may play an important role in expression of individual crystal proteins in B. thuringiensis without doubt.