• Title/Summary/Keyword: ITS2-rDNA region

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Flowers in Islands and Mountains of Korea

  • Hyun, Se-Hee;Kim, Ha-Kun;Lee, Jong-Soo
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.44-44
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    • 2014
  • Yeasts were isolated from wild flowers of some islands and mountains such as Jeju-do, Ulleungdo, Yokjido, Seonyudo and Gyejoksan, Oseosan, Beakamsan and Deogyusan in Korea and were identified by comparison of nucleotide sequences for PCR-amplified D1/D2 region of 26S rDNA or internal transcribed pacer(ITS) 1 and 2 including 5.8S rDNA using BLAST. Seventy two yeast strains of two hundred eighty nine species were isolated from wild flowers in islands and mountains, Korea. Among them, Cryptococcus species were isolated the most dominantly, and Metschnikowia reukaufii were also isolated thirty species, 10.3% of total strains. Twenty-three species including Cryptococcus aureus were overlapped between yeast strains of the islands and mountains. Some physiological functionality of the culture broth and cell-free extracts from two hundred eighty nine yeast strains were determined. The supernatant of Candida sp. 78-J-2 showed antioxidant activity of 22.5%, and supernatant of Metschnilowia reukaufii SY44-6 showed anti-gout xanthine oxidase inhibitory activity of 49.6% and whitening tyrosinase inhibitory activity of 38.4%, respectively.

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DNA Structural Perturbation Induced by the CPI-Derived DNA Interstrand Cross-linker : Molecular Mechanisms for the Sequence Specific Recognition

  • Park, Hyun-Ju
    • Archives of Pharmacal Research
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    • v.24 no.5
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    • pp.455-465
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    • 2001
  • The highly potent cytotoxic DNA-DNA cross-linker consists of two cyclopropa[c]pyrrolo[3,4-3]indol-4(5H)-ones insoles [(+)-CPI-I] joined by a bisamido pyrrole (abbreviated to "Pyrrole"). The Pyrrole is a synthetic analog of Bizelesin, which is currently in phase II clinical trials due to its excellent in vivo antitumor activity. The Pyrrole has 10 times more potent cytotoxicity than Bizelesin and mostly form DNA-DNA interstrand cross-links through the N3 of adenines spaced 7 bp apart. The Pyrrole requires a centrally positioned GC base pair for high cross-linking reactivity (i.e., $5^1$-T$AT_2$A*-$3^1$), while Bizelesin prefers purely AT-rich sequences (i.e., $5^1$-T$AT_4$A*-$3^1$, where /(equation omitted) represents the cross-strand adenine alkylation and A* represents an adenine alkylation) (Park et al., 1996). In this study, the high-field $^1$H-NMR and rMD studies are conducted on the 1 1-mer DNA duplex adduct of the Pyrrole where the 5′(equation omitted)TAGTTA*-3′sequence is cross-linked by the drug. A severe structural perturbation is observed in the intervening sequences of cross-linking site, while a normal B-DNA structure is maintained in the region next to the drug-modified adenines. Based upon these observations, we propose that the interplay between the bisamido pyrrole unit of the drug and central C/C base pair (hydrogen-bonding interactions) is involved in the process of cross-linking reaction, and sequence specificity is the outcome of those interactions. This study suggests a mechanism for the sequence specific cross-linking reaction of the Pyrrole, and provides a further insight to develop new DNA sequence selective and distortive cross-linking agents.

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Identification of Aspergillus Strain with Antifungal Activity Against Phytophthora Species

  • KANG SUNG WOO;HONG SUK IN;KIM SEUNG WOOK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.227-233
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    • 2005
  • Fungal strain CGF was isolated from the soil of ChungNam Province, South Korea. Based on the 28S rDNA sequence analysis and the sequence of the internal transcribed spacer (ITS) region of ribosomal DNA, together with morphological and cultural characteristics, this strain was identified as Aspergillus sclerotiorum and renamed Aspergillus sclerotiorum CGF. This is the first strain of Aspergillus sclerotiorum identified in Korea. When the antifungal activity of A. sclerotiorum CGF was evaluated, among the phytopathogenic fungi, mycelial growth of only Phytophthora species was inhibited. Oermination of P. capsid zoospore was also inhibited. The bioactive compound of A. sclerotiorum CGF was highly thermo- and pH-stable.

Genomic Organization and Characterization of the Promoter Region of Bovine ADRP (Adipocyte Different Related Protein) Gene (소 Adipocyte Differentiation Related Protein (ADRP) 유전자의 Genomic Organization 및 Promoter Region의 특성 규명)

  • Jang, Y. S.;Yoon, D. H.;Kim, T. H.;Cheong, I. C.;Jo, J. K.
    • Journal of Animal Science and Technology
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    • v.45 no.2
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    • pp.169-182
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    • 2003
  • To understand the structure and regulation of bovine ADRP (Adipocyte Differentiation Related Protein) gene, we have isolated the genomic clone of bovine ADRP and determined its sequence. A genomic Southern blot analysis confirmed that ADRP gene is present as a single copy in bovine genome and the ADRP gene spans 12 kb. Bovine ADRP genomic clone, HwADRPg-1, had 8 exons and 7 introns, and all splicing sites conformed to the GT/AG rule with the exon-intron boundaries located exactly. Analysis of the upstream 649 bp of the sequence of HwADRPg-1 showed that it does not contain any canonical TATAA boxes; however Sp1 binding sites and CAAT boxes are found. The promoter contained potential binding sites for AP-1, AP-2 and several putative transcription factor binding sites. The 5'-flanking region of HwADRPg-1 contained muscle specific transcription activator Myo G and C/EBP (CCAAT/ enhancer binding protein) recognizing site. These results suppose that the Myo G transcription activator regulate the transcription of bovine ADRP gene in muscular tissue and its transcriptional activity was triggered by degree of muscular development. Our results provide the necessary analysis for other flanking sequences are needed in addition to the proximal cis elements of this promoter to confer adipocyte differentiation-dependent or growth-dependent transcriptional control.

Diversity of Endophytes Isolated from Thuja koraiensis Nakai in the Korean Peninsula (눈측백(Thuja koraiensis Nakai)에서 분리한 내생균의 다양성)

  • Eo, Ju-Kyeong;Lee, Bong-Hyung;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.44 no.2
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    • pp.113-117
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    • 2016
  • The biodiversity of endophytic fungi on Thuja koraiensis in Mt. Hwaak, Seorak, and Hambaek, Korea was investigated. For the 202 isolates collected from the host trees, internal transcribed spacer rDNA region sequences-based analysis identified 32 taxa; 61.5% of the isolates belonged to Dothideomycetes, 27.0% belonged to Sordariomycetes, and 11.5% belonged to Leotiomycetes. This composition rate is somewhat different from that reported in previous studies for endophytic fungi inhabiting trees of the family Pinaceae. In particular, Phyllosticta spinarum in Dothideomycetes is a dominant species among the diverse endophytes of T. koraiensis. Therefore, further critical research is required for this species.

Molecular Cloning and Recombinant Expression of the Long Form of Leptin Receptor (Ob-Rb) cDNA as Isolated from Rat Spleen

  • Ju, Sung-Kyu;Park, Jung-Hyun;Na, Shin-Young;You, Kwan-Hee;Kim, Kil-Lyong;Lee, Myung-Kyu
    • BMB Reports
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    • v.34 no.2
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    • pp.156-165
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    • 2001
  • Leptin is a circulating non-glycosylated protein that is mainly produced in adipocytes. Leptin acts in the brain to regulate food intake and energy expenditure. Previously we reported our success in the isolation of a partial cDNA of the long form of the leptin receptor, OB-Rb, from rat spleen, and showed that leptin might also play a role in peripheral immune organs. In the present study, for the first time, the complete coding region of OB-Rb cDNA was cloned from rat splenocytes, and its nucleotide sequence was determined. The cDNA was then further expressed in E. coli and mammalian cells, thereby confirming the functional integrity of this receptor. Prokaryotically overexpressed OB-R protein was then used as an immunizing antigen in BALE/c mice to produce leptin receptor-specific antibodies. By using them, we confirmed the cell surface expression of OB-Rb in transfected CHO cells. It is our belief that the reagents, as produced in this study, will be of great use in further studies of the biological role of rat leptin.

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Characterization and Regulation of the Gene Encoding Monothiol Glutaredoxin 3 in the Fission Yeast Schizosaccharomyces pombe

  • Moon, Jeong-Su;Lim, Hye-Won;Park, Eun-Hee;Lim, Chang-Jin
    • Molecules and Cells
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    • v.20 no.1
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    • pp.74-82
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    • 2005
  • Glutaredoxins (Grxs) are thioloxidoreductases which are required for maintaining thiol/disulfide equilibrium in living cells. The Grx3 gene, which encodes one of the three monothiol Grxs in the fission yeast Schizosaccharomyces pombe, was characterized, and its transcriptional regulation studied. Genomic DNA encoding Grx3 was isolated by PCR, and a plasmid pTT3 carrying this DNA was produced. The DNA sequence has 1,267 bp, which would encode a monothiol Grx of 166 amino acids with a molecular mass of 18.3 kDa. The putative protein has 27% homology with Grx5, and contains many hydrophobic amino acid residues in its N-terminal region. S. pombe cells harboring pTT3 had increased Grx activity and enhanced survival on minimal medium plates containing aluminum (5 mM), BSO (0.05 mM), menadione (0.01 mM) or cadmium (0.2 mM). The 568 bp upstream region of Grx3 was fused into the promoterless b-galactosidase gene of the shuttle vector YEp367R to generate fusion plasmid pMJS10. Potassium chloride (KCl) and metals including aluminum and cadmium enhanced the synthesis of ${\beta}$-galactosidase from the fusion gene. The synthesis of ${\beta}$-galactosidase was also enhanced, in a Pap1-dependent manner, by fermentable carbon sources such as glucose (at low concentrations) and sucrose, but not by non-fermentable carbon sources such as ethanol and acetate. Grx3 mRNA increased in response to treatment with BSO. These observations indicate that S. pombe Grx3 is involved in the response to stress, and is regulated by stress.

Morphological and Multigene Sequence Characteristics of Talaromyces variabilis Isolated from Soil in Korea

  • Adhikari, Mahesh;Kim, Sang Woo;Lee, Hyang Burm;Lee, Youn Su
    • The Korean Journal of Mycology
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    • v.49 no.1
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    • pp.11-19
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    • 2021
  • In 2017, Talaromyces variabilis was isolated during a survey of fungal diversity in field soils in Korea. This isolate was described based on its morphological and molecular characteristics and it was identified molecularly using the partial 18S-ITS1-5.8S-ITS2-28S rDNA region and calmodulin (CaM)-encoding gene sequence data. Thus, this study reported morphological and multigene sequence characterization of T. variabilis.

Isolation and Identification of Yeasts from Wild Flowers of Daejeon City and Chungcheongnam-do in Korea (대전과 충남 지역에서 채취한 야생화로부터 효모의 분리 및 동정)

  • Min, Jin-Hong;Hyun, Se-Hee;Kang, Min-Gu;Lee, Hyang-Beom;Kim, Chang-Mu;Kim, Ha-Kun;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.40 no.3
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    • pp.141-144
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    • 2012
  • Total 50 species, 80 strains: Eighty strains of yeasts were isolated from wild flowers of Daejeon city and its surroundings in Korea. Isolated yeasts were identified by comparison of their PCR-amplified 18s rDNA V3 region or internal transcribed spacer-2 (ITS-2) genes with BLAST database. Among them, 32 species, 54 strains of yeasts were isolated and identified from wild flowers of Daejeon city, and Cryptococcus species were dominant yeasts. Furthermore, 18 species, 26 strains of yeasts were isolated and identified from wild flowers of Geumsan-gun and Nonsan-gun, Chungcheongnam-do in Korea, and also Candida species and Pseudozyma species were dominant yeasts.

Isolation and Identification of Yeasts from Wild Flowers in Gyejoksan, Oseosan and Beakamsan of Korea (대전 계족산과 충남 오서산 및 전북 백암산 주위 야생화들로부터 효모의 분리 및 동정)

  • Min, Jin-Hong;Ryu, Jin-Ju;Kim, Ha-Kun;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.41 no.1
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    • pp.47-51
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    • 2013
  • Yeasts isolated from wild flowers of Gyejoksan in Daejeon city, Oseosan in Chungchungnamdo, and Baekamsan in Jeollabukdo, Korea were identified by comparison of nucleotide sequences for PCR-amplified D1/D2 region of 26S rDNA or internal transcribed spacer (ITS) 1 and 2 including 5.8S rDNA using BLAST. Twelve yeast strains of ten species and seventeen yeast strains of ten species were isolated from wild flowers of Gyejoksan and Oseosan, respectively. And thirty seven yeast strains of twenty four species were isolated from wild flowers of Baekamsan. Total thirty four yeast species were isolated from three different sample collection areas, but only nine species were overlapped from the at least two different sampling areas: Cryptococcus sp., Cryptococcus aureus, Cryptococcus flavescens, Cryptococcus flavus, Metschnikowia sp., Pseudozyma aphidis, Rhodotorula glutinis, Sporobolomyces carnicolor, and Sporobolomyces ruberrimus. Among them only Cryptococcus aureus was occurred from all three different collection sites. Other twenty five species were restricted to specific collection site suggesting that each area has distinctive yeast flora.