• Title/Summary/Keyword: 25S rDNA

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Genetic Relationships of Coprinus spp. on the Basis of Sequences in ITS II Region (먹물버섯류(Coprinus spp.)의 ITS II 영역 염기서열에 의한 유연관계 분석)

  • Park, Dong-Suk;Go, Seung-Joo;Kim, Yang-Sup;Seok, Soon-Ja;Song, Jae-Kyeong;Yeo, Yun-Soo;Ryu, Jin-Chang;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.27-31
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    • 1999
  • The internal transcribed spacer II regions (ITS II) of the ribosomal DNA gene repeat from Coprinus spp. were amplified using polymerase chain reaction (PCR) and sequenced. Sequences from 11 species including Coprinus comatus, C. atramentarius, C. micaceus, C. lagopus, C. cinereus, C. rhizophorus, C. flocculosus, C. radians, and C. echinosporus were compared. The spacer regions of them were $253{\sim}275$ nucleotide in length and partially contained 5.8S and 25S. The reciprocal homologies of each ITS II sequence among these strains were in the range of $50.6{\sim}100%$. According to the analysis of ITS II sequences, Coprinus spp. were classified into three clusters. Cluster I consisted of Coprinus lagopus, C. cinereus, C. echinosporus, C. rhizophorus, C. niveus, and C. atramentarius. Cluster II comprised C. micaceus, C. flocculosus, C. radians, and C. disseminatus. On the other hand C. comatus is in Cluster III even though this species is belonging to the section Coprinus in morphological aspect. These results suggest that Coprinus comatus, which was considered as a type species of the genus Coprinus in morphological classification, gives a doubt of monophyletic evolution and is assumed to be paraphyletic or polyphyletic.

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Characterization of Bovine Lymphocyte Antigen DRB3 exon2 Gene of Korean Native Cattle (한우의 BoLA DRB3 exon2 유전자의 특성)

  • Kang, Ho Bum;Ryoo, Seung Heui;Lee, Sang Hoon;Jeon, Byung Soon;Sang, Byung Chan
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.79-88
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    • 1998
  • This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.

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Isolation and Identification of Biofilm-Forming Marine Bacteria on Glass Surfaces in Dae-Ho Dike, Korea

  • Kwon, Kae-Kyoung;Lee, Hyun-Sang;Jung, Sung-Young;Yim, Joung-Han;Lee, Jung-Hyun;Lee, Hong-Kum
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.260-266
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    • 2002
  • Bacterial strains were isolated from biofilms formed on glass slides submerged in seawater in Dae-Ho Dike. Eight strains showing fast attaching ability were selected and identified. Their exopolysaccharide (EPS)-producing ability and EPS properties were characterized. Based on Microlog System, 4 among the 8 strains were identified as Micrococcus luteus and the rest were Bacillus thuringiensis, Bacillus megaterium,, Staphylococcus saprophyticus and Agrobacterium vitis. A, vitis was reidentified as Sulfitobacter pontiacus based on 16S rDNA sequence data. The amount of water-soluble EPS produced by the 8 strains ranged from 0.114 to 1.329 g$.$l$\^$-1/ and the productivity was negatively correlated with the cell biomass. The molecular weight of the produced EPS ranged from 0.38 to 25.19$\times$10$\^$4/ Da. Glucose and galactose were ubiquitous sugar components. Mannose, ribose, and xylose were also major sugar components. The molecular weight and composition of the EPS showed strain-specific variation.

Diversity of Cultured and Uncultured Bacteria in the Gut of Olive Flounder Paralichthys olivaceus (넙치(Paralichthys olivaceus) 장관의 배양 및 비배양 방법에 의한 세균의 다양성)

  • Kim, Ahran;Kim, Do-Hyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.4
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    • pp.447-453
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    • 2015
  • We determined the optimal culture conditions for obtaining the maximum number of intestinal bacteria from the olive flounder Paralichthys olivaceus, and studied bacterial diversity using both culture-dependent and culture-independent methods. Using six culture conditions, mean bacterial numbers were greater than $10^6$ per gram of gut mucus, regardless of the medium. However, the bacterial diversity, based on colony morphology, appeared much higher on Marine agar (MA) and Zobell 2216 agar than on other media. We found eight and 17 cultured bacterial phylotypes with 99% minimum similarity in gut mucus grown on MA and tryptic soy agar, respectively. Furthermore, we used genomic DNA extracted from gut mucus to generate 78 random clones, which were grouped into 25 phylotypes. Of these, six were affiliated with Firmicutes, Actinobacteria, and Verrucomicrobia, and were not found using our culture-dependent methods. Consequently, we believe that Marine agar and Zobell 2216 agar are optimal media for culturing diverse intestinal microbes; we also discovered several novel sequences not previously recognized as part of the gut microbiota of olive flounder.

Studies on the Development of a Microbial Cryoprotectant Formulation Using a W/O/W Multiple Emulsion System

  • Bae, Eun-Kyung;Cho, Young-Hee;Park, Ji-Yong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.673-679
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    • 2004
  • A microbial cryoprotectant formulation using a W/O/W multiple emulsion system was developed. The psychrotolerant microorganism, B4, isolated from soil in South Korea, was observed by the drop freezing method, in which the microorganism sample inhibited ice nucleation activity. The antifreeze activity was eliminated when the microorganism sample was treated with protease, indicating that the antifreeze activity was due to the presence of antifreeze protein. The result of the l6S rDNA sequencing indicated the B4 strain was most closely related to a species of the genus Bacillus. Culture broth of B4 strain (Bacillus sp.) and rapeseed oil containing 1 % polyglycerine polyricinolate (PGPR) were used as core and wall material, respectively. The most stable W/O emulsion was prepared at a core/oil ratio of 1:2. The highest W/O/W emulsion stability was achieved when the primary emulsion to external aqueous phase containing 0.5% caster oil polyoxyethylene ether $(COG25^{TM})$ ratio was 1:1. Microcrystalline cellulose showed better W/O/W emulsion stability than other polymer types. The viability of cells in a W/O/W emulsion was higher than free cells during storage at $37^\circ{C}$. An acidic pH and UV exposure decreased the viability of free cells, but cells in W/O/W emulsion were more stable under these conditions.

Isolation and Characterization of a Protease-Producing Halophilic Vibrio sp. (단백질 분해효소를 생산하는 호염성 미생물 Vibrio의 분리 및 특성)

  • Um, Ki-Bum;Yoon, Sun-Jin;Lee, Jae-Kyoung;Lee, Jae-Hag;Lee, Soon-Youl
    • The Korean Journal of Food And Nutrition
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    • v.20 no.2
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    • pp.108-113
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    • 2007
  • In this study, a halophilic protease-producing bacterium was isolated from the west seaside mud flats of Korea. The 16S rDNA nucleotide sequences of the isolate showed 99.5% sequence homology with those of Vibrio vulnificus and Vibrio fluvialis; therefore, the isolate was named Vibrio sp. YH-127. Gram staining and the carbohydrate metabolism test results supported the isolate as one from the Vibrio family. Optimum condition for the cell growth and for the protease activity were obtained when the isolate was cultured at 25$^{\circ}C$ and pH 7.0, with the salt concentration of the medium similar to that of sea water. Finally, the addition of Mg$^{++}$ ions into medium increases protease activity suggesting that the protease produced by the isolate was a metalloprotease.

Characterization of a Nitrogen Fixing Bacteria Mycobacterium hominis sp. AKC-10 Isolated from the Wetland (습지에서 분리한 질소고정 세균인 Mycobacterium hominis sp. AKC-10의 특성)

  • Hong, Sun-Hwa;Shin, Ki-Chul;Lee, Eun-Young
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.302-307
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    • 2010
  • Nitrogen is an element need to grow plants growth. Plants take up nitrogen in the form of nitrate or ammonium. Most of plants absorb nitrogen source as fertilizers. But from 50 to 70% of fertilizers applied were washed away. This study was conducted to isolate free-living nitrogen fixing bacteria from reed and to examine its beneficial traits for developing sustainable biofertilizers. Enriched consortium obtained from a reed in Ansan was developed for the fixing of nitrogen. Nitrogen fixing bacteria isolated from an enriched culture in Congo Red Medium was analyzed by 16s rDNA sequencing. AKC-10 was isolated and shown to have excellent nitrogen fixing ability. The optimum conditions of nitrogen fixing ability were $25^{\circ}C$ ($237.50{\pm}39.65\;nmole{\cdot}mg-protein^{-1}{\cdot}h^{-1}$ and pH 7 ($168.335{\pm}12.84$ nmole/hr mg-protein). It was identified as Microbacterium hominis [(AKC-10 (similarity : 99%)]. This strain was had to IAA (indole-3-acetic acid) productivity and ACC(1-aminocyclopropane-1-carboxylic acid) deaminase activity. Therefore, Microbacterium hominis AKC-10 stimulated plant development in the soil, enhancing the efficiency of remediation.

Development of Thermostable Fusant, CHY1612 for Lignocellulosic Simultaneous Saccharification and Fermentation (섬유질계 동시당화발효를 위한 내열성 융합 효모, Kluyveromyces marxianus CHY1612의 개발)

  • Kang, Hyun-Woo;Kim, Yule;Park, Ju-Yong;Min, Ji-Ho;Choi, Gi-Wook
    • KSBB Journal
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    • v.25 no.6
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    • pp.565-571
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    • 2010
  • To develop thermostable ethanol fermentative yeast strain for lignocellulosic simultaneous saccharification and fermentation, high ethanol producing yeast, Saccharomyces cerevisiae CHY1012 and thermostable yeast, Kluyveromyces marxianus CHY1703 were fused by protoplast fusion. The thermostable fusant, CHY1612 was identified as a Kluyveromyces marxianus by phenotypic and physiological characteristics, as well as molecular analysis based on the D1/D2 domains of the large subunit (26S) rDNA gene and the internally transcribed spacer (ITS) 1 + 2 regions. For lignocellulosic ethanol production, AFEX pretreated barley straw at $150^{\circ}C$ for 90 min was used in a simultaneous saccharification and fermentation (SSF) process using thermotolerant CHY1612. The SSF from 16% pretreated barley straw at $43^{\circ}C$ gave a saccharification ratio of 90.5%, a final ethanol concentration of 38.5 g/L, and a theoretical yield of 91.2%. These results show that K. marxianus CHY1612 has potential for lignocellulosic ethanol production through simultaneous saccharification and fermentation with further development of process.

A New Report of Biscogniauxia petrensis Isolated from Mosquitoes in Korea

  • Das, Kallol;Kim, Joung-Ho;Choi, Kwang-Shik;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.48 no.2
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    • pp.87-93
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    • 2020
  • A fungal strain designated KNU-WDM2A2 was isolated from mosquitoes in Gimcheon, Korea. The pure culture was transferred to potato dextrose agar (PDA) and synthetic nutrient agar (SNA) media and attained a diameter of 90 mm after 10 days of incubation at 25℃. The colonies were whitish to light pink and cottony to wooly, with an abundant production of aerial mycelia. The strain produced hyaline to slightly yellowish conidiophores that were rough-walled and branched, with conidiogenous cells arising terminally or laterally. Conidia were unicellular, hyaline to light brown, smooth, and oval or ovoid to clavate, with a size of 4.1-6.9×2.5-3.3 ㎛ (n=65). A phylogenetic analysis was conducted using the internal transcribed spacer (ITS) regions and 28S rDNA of large subunit (LSU) sequences, to support the cultural and morphological characteristics. The KNU-WDM2A2 strain was identified here as Biscogniauxia petrensis, new to Korea.

Morphological and Phylogenetic Analysis of a New Record of Paraconiothyrium kelleni from Soil in Korea

  • Mukesh Kumar Yadav;Kallol Das;Jung-Joo Ryu;Seong-Keun Lim;Jin-Sil Choi;Seung-Yeol Lee;Hee-Young Jung
    • The Korean Journal of Mycology
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    • v.51 no.2
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    • pp.101-109
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    • 2023
  • A fungal strain designated KNUF-21-66Q1 was isolated from soil in Chungcheongbuk Province, Korea. Moderate growth of colonies was observed on potato dextrose agar, oatmeal agar (OA), malt extract agar, and cornmeal agar media at 25℃, and the detailed morphology was examined on OA medium. The colonies on OA medium were flat, had entire margin, hyaline, and yellow concentric rings in 3-4 weeks. Conidiomata were pycnidial, solitary or clustered, globose to subglobose, black-brown, and 300-500 ㎛ in diameter. Conidiogenous cells were smooth, hyaline, globose to ampulliform, and 6.0-9.0×3.0-6.0 ㎛ in size (n=15). Conidia were hyaline to pale brown, slightly golden, obovoid to slightly ellipsoidal, smooth, guttulate, and 3.0-4.7×2.1-3.3 ㎛ in size (n=100). The strain was confirmed based on phylogenetic analysis using internal transcribed spacer regions, the partial 28S rDNA of large subunit, and β-tubulin gene sequences. The morphological observations and phylogenetic analysis revealed that the strain KNUF-21-66Q1 was similar to the previously described Paraconiothyrium kelleni. To our knowledge, this is the first report of P. kelleni in Korea.