• Title/Summary/Keyword: 5.8S rRNA

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Intrageneric Relationships of Trichoderma Based on Internal Transcribed Spacers and 5.8S rDNA Nucleotide Sequences

  • Kim, Gi-Young;Lee, Goang-Jae;Ha, Myung-Gyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.28 no.1
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    • pp.11-16
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    • 2000
  • The nucleotide sequences of the internal transcribed spacer (ITS) regions of the ribosomal DNA including the 5.8S ribosomal RNA gene (rDNA) have been determined for 11 species in order to analyze their intrageneric relationships. The total length of these sequences ranged from 530 nucleotides for Trichoderma reesei KCTC 1286 to 553 nucleotide for Trichoderma koningii IAM 12534. Generally speaking, the length of ITS1 region was about 30 nucleotides longer than that of the ITS2 region. Also, the sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites were also found. Thus, a neighbor-joining tree was constructed using the full sequence data of the ITS regions and the 5.8S rDNA. The Trichoderma genus used to be grouped on the basis of the morphological features and especially the shape of phialides needs to be reexamined. The phylogenetic tree displayed the presence of monophylogeny in the species of Trichoderma. Therefore, it was difficult to distinguish the intrageneric relationships in the Trichoderma genus.

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Clonorchis sinensis tropomyosin: Cloning and sequence of partial cDNA amplified by PCR (간흡충 tropomyosin: PCR로 일부분 증폭된 cDNA의 cloning 및 염기서열)

  • 홍성종
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.285-292
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    • 1993
  • C. sinensis total RMh was containing large amount of 185 rRNA but little 285 rRNA. The size of the double-stranded cDNA synthesized from poly $(A)^{+}$ mRNA was 0.4-4.2 kb long with tapering unto 9.5 kb. Degenerated oligonucleotides (as 2 sense and 3 antisense Primers) were designed on the conserved regions of the known tropomyosin amino acid sequences. From one out of the PCR amplifications using total CDNA and matrix of primers, a specific gene product, 580 bp in size, was produced. Upon Southern hybridization of the PCR products with Schistosomn mnnsoni tropomyosin (SMTM) CDNA, only one signal appeared at the band of 580 bp product. This 580 bp product was considered to encode C. sinensis tropomyosin (CSTM) and cloned in pGEM-3Zf(-) for DNA sequencing. CSTM cDNA was 575 bp containing one open reading frame of 191 predicted amino acids, which revealed 86.3% homology with SMTM and 51.1% with rrichostronsylur coeubnlormis tropomyosin. CSTM cDNA obtained will serve as a probe in the studies of molecular cloning of CSTM.

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Quantitative Analysis of Bacillus amyloliquefaciens GR4-5 in Soil (Bacillus amyloliquefaciens GR4-5 균주의 토양 내 정량 분석)

  • Kim, Dayeon;Kim, Byung-Yong;Ahn, Jae-Hyung;Weon, Hang-Yeon;Kim, Sung-Il;Kim, Wan-Gyu;Song, Jaekyeong
    • Korean Journal of Organic Agriculture
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    • v.23 no.4
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    • pp.847-858
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    • 2015
  • Bacillus amyloliquefaciens GR4-5 was isolated from the rhizosphere soil of Korean ginseng and displayed broad-spectrum suppression of ginseng root rot pathogens. The survivability of B. amyloliquefaciens GR4-5 in soil was investigated under three different conditions; indoor, outdoor - of which soil was put in 14 mL tube after treatment - and field environments. Soil samples were collected over a four-week period from three experimental designs, and assessed for 16S rRNA gene copy number by quantitative polymerase chain reaction (qPCR). In outdoor condition, the 16S rRNA gene copy number of Bacillus spp. was 8.35 log copies g $soil^{-1}$ immediately after the GR4-5 treatment. Two weeks later, the 16S rRNA gene copy number of Bacillus spp. (6.70 log copies g $soil^{-1}$) was similar to that of the control (6.38 log copies g $soil^{-1}$). In indoor condition, the 16S rRNA gene copy number of Bacillus spp. maintained in a certain level for a longer period than those in outdoor and field. The 16S rRNA gene copy number of Bacillus spp. in field experiment was reduced faster than that of outdoor condition. Our results show that B. amyloliquefaciens GR4-5 can survive in bulk soil for 1 week, indicating its potential use as a biocontrol agent following 7 day application intervals. This study presents that outdoor microcosm system design could be a useful method to assess easily the survivability of beneficial microorganisms.

Cloning and Characterization of Mannanase Gene from Bacillus subtilis WL-8 (Bacillus subtilis WL-8의 Mannanase 유전자 클로닝과 특성분석)

  • Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.207-212
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    • 2010
  • A bacterium producing the extracellular mannanase was isolated from Korean soybean paste. The isolate WL-8 has been identified as Bacillus subtilis on the basis on its 16S rRNA sequence, morphology and biochemical properties. The mannanase productivity of strain WL-8 was increased in LB broth by addition of wheat bran. The maximum mannanase productivity was reached to approximately 20 U/ml in LB medium supplemented with 6% wheat bran. A gene encoding the mannanase of WL-8 was cloned into Escherichia coli and its nucleotide sequence was subsequently determined. The mannanase gene consisted of 1,086 nucleotides encoding a polypeptide of 362 amino acid residues. The deduced amino acid sequence was highly homologous with those of several mannanases from B. subtilis belonging to GH family 26. Reaction temperature and pH profiles were investigated using the culture filtrate and cell-free extract of the recombinant E. coli carrying a WL-8 mannanase gene, respectively. Optimal conditions for the two fractions occurred at pH 5.5 and $60^{\circ}C$. The cell-free extract showed higher mannanase activity than the culture filtrate at above $60^{\circ}C$.

Microbial Diversity of the Trichloroethylene Contaminated Groundwater Treatment System and Characterization of Pseudomonas sp. DHC8 (Trichloroethylene으로 오염된 지하수 제거공정의 미생물 다양성 및 분리균주 Pseudomonas sp. DHC8의 특성)

  • Nam, Ji-Hyun;Shin, Ji-Hye;Kwon, Kiwook;Bae, Wookeun;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.336-342
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    • 2013
  • Trichloroethylene (TCE) is a widely used substance in commercial and industrial applications, yet it must be removed from the contaminated soil and groundwater environment due to its toxic and carcinogenic nature. We investigated bacterial community structure, dominant bacterial strain, and removal efficiency in a TCE contaminated groundwater treatment system using immobilized carrier. The microbial diversity was determined by the nucleotide sequences of 16S rRNA gene library. The major bacterial population of the contaminated groundwater treatment system was belonging to BTEX degradation bacteria. The bacterial community consisted mainly of one genus of Pseudomonas (Pseudomonas putida group). The domination of Pseudomonas putida group may be caused by high concentration of toluene and TCE. Furthermore, we isolated a toluene and TCE degrading bacterium, named Pseudomonas sp. DHC8, from the immobilized carrier in bioreactor which was designed to remove TCE from the contaminated ground water. Based on the results of morphological and physiological characteristics, and 16S rRNA gene sequence analysis, strain DHC8 was identified as a member of Pseudomonas putida group. When TCE (0.83 mg/L) and toluene (60.61 mg/L) were degraded by this strain, removal efficiencies were 72.3% and 100% for 12.5 h, respectively. Toluene removal rate was 2.89 ${\mu}mol/g$-DCW/h and TCE removal rate was 0.02 ${\mu}mol/g$-DCW/h. These findings will be helpful for maintaining maximum TCE removal efficiency of a reactor for bioremediation of TCE.

Phylogenetic Relationships among Some Bumblebees (Hymenoptera: Apidae) Common in Korea Inferred from Mitochondrial 16S rRNA Sequences

  • Yoon, Hyung-Joo;Lee, Myeong-Lyeol;Kim, Sam-Eun;Lee, Sang-Beom;Kim, Iksoo;Bae, Jin-Sik;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.1
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    • pp.51-59
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    • 2004
  • Phylogenetic relationships were analyzed among bumblebees using a portion of mitochondrial (mt) 16S ribosomal RNA (16S rRNA). Eight species of true bumblebees and one species of cuckoo bumblebee (Bombini, Apidae), collected from Korea were included in the analysis. Also, one species of true bumblebee imported from several foreign countries for pollination was included. The length of mt 16S rRNA sequence ranged from 496 bp to 508 bp and sequence divergence ranged from 1.4% (7 bp) to 15.49% (77bp). As expected, a high A+T content was observed (78.5% on average). According to the phylogeny tree derived from parsimony and maximum likelihood analysis, a monphyletic Bombus species, excluding a single cuckoo bumblebee, Psithyrus coreanus, was obtained, but the bootstrap estimate at the node supporting the monophyletic group was very weak (40% or 46%), suggesting a very close relationship of the cuckoo bumblebee to the true bumblebee. Within Bombus species belonging to identical subgenera subgeneric specific clustering was formed with high bootstrap values, implying validity of the subgeneric names of each species: Pyrobombus for B. ardens and B. modeatus; Megabombus for B. consobrinus wittenburgi and B. koreanus; and Bombus s. str. for B. ignitus, B. hypocrita sapporoensis, and B. terrestris.

Taxonomic characteristics of novel Flavobacteriumsp. B1 from a freshwater pond

  • Bae, Young-Min
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.5
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    • pp.605-613
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    • 2022
  • The genus Flavobacterium, type genus of the family Flavobacteriaceae and a member of the phylum Bacteriodetes includes gram-negative and yellow-pigmented rods. Those bacteria have been isolated from various environments of the earth. A yellow-pigmented, gram-negative rod was isolated from a pond in the campus of the Changwon University, Changwon, Kyeongnam and designated as strain B1. Strain B1 was further analyzed physiologically, biochemically and phylogenetically, and concluded to be a member of genus Flavobacterium. BLAST search of the 16S rRNA gene sequence of strain B1 shows homology no higher than 99.0% with those sequences of other bacteria. The major fatty acids of strain B1 are iso-C15:0 (19.6%), summed feature 3(C16:1 ω7c and/or C16:1 ω6c, 16.1%), iso-C17:0 3OH(10.2%), iso-C15:0 3OH(8.4%) and iso-C15:1 G(6.6%) showing significant differences in fatty acid compositions between strain B1 and the other known Flavobacterium species. DNA sequence of 16S rRNA gene of strain B1 was deposited in genbank under accession number OP060681.

Methanogenic Archaeal Census of Ruminal Microbiomes (반추위 마이크로바이옴 내 메탄생성고세균 조사)

  • Lee, Seul;Baek, Youlchang;Lee, Jinwook;Kim, Minseok
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.7
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    • pp.312-320
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    • 2020
  • The objective of the study was to undertake a phylogenetic diversity census of ruminal archaea based on a meta-analysis of 16S rRNA gene sequences that were publicly available in the Ribosomal Database Project. A total of 8,416 sequences were retrieved from the Ribosomal Database Project (release 11, update 5) and included in the construction of a taxonomy tree. Species-level operational taxonomic units (OTUs) were analyzed at a 97% sequence similarity by using the QIIME program. Of the 8,416 sequences, 8,412 were classified into one of three phyla; however, the remaining four sequences could not be classified into a known phylum. The Euryarchaeota phylum was predominant and accounted for 99.8% of the archaeal sequences examined. Among the Euryarchaeota, 65.4% were assigned to Methanobrevibacter, followed by Methanosphaera (10.4%), Methanomassillicoccus (10.4%), Methanomicrobium (7.9%), Methanobacterium (1.9%), Methanimicrococcus (0.5%), Methanosarcina (0.1%), and Methanoculleus (0.1%). The 7,544 sequences that had been trimmed to the V2 and V3 regions clustered into 493 OTUs. Only 17 of those 493 OTUs were dominant groups and accounted for more than 1% of the 7,544 sequences. These results can help guide future research into the dominant ruminal methanogens that significantly contribute to methane emissions from ruminants, research that may lead to the development of anti-methanogenic compounds that inhibit these methanogens regardless of diet or animal species.

The effects of the mulberry and silkworm intake on androgen receptor mRNA and myogenic regulatory factors expression of rats muscle for resistance exercise (오디와 누에 섭취가 rats의 저항성 운동에 따른 androgen receptor mRNA와 myogenic regulatory factors의 발현에 미치는 영향)

  • Yang, Sung Jun;Kim, Chang Yong;Lee, Jo Byoung;Kang, Sung Sun;Lee, Jong Jin
    • Journal of Sericultural and Entomological Science
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    • v.51 no.2
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    • pp.99-106
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    • 2013
  • The purpose of this study is to investigate the effects of supplementation of mulberry powder, mulberry extract and silkworm powder during the 8 weeks of resistance exercise on Androgen receptor(AR) mRNA and Myogenic regulatory factors(MRFs) expression of rats muscle. Fifty males, Sprague-Dawley rat, were randomly divided into 5 groups: CON(control group, n = 10), REG(resistance exercise group, n = 10), MP REG(mulberry powder intake and resistance exercise group, n = 10), ME REG(mulberry extract intake and resistance exercise group, n = 10) and SP REG(silkworm powder intake and resistance exercise group, n = 10). After climbing the ladder without weights during the 1 week of adaptation period, the rats in the resistance exercise group were trained to climb a 0.98-m vertical(80 degree incline) ladder with weights in their tail during 7 weeks(10 times each day, 2 days per week). After exercise, the skeletal muscle was extracted from the flexor hallucis longus. After separating the total ribonucleic acid (RNA) of each group, quantitative polymerase chain reaction was used to analyze RNA quantitatively. AR mRNA and MRFs expression revealed that all of the treated groups had significantly difference. AR mRNA expression increased in ME REG $6.24{\pm}1.85$ and SP REG $9.68{\pm}0.28$ fold compared to CON. Myod mRNA expression increased in MP REG $6.04{\pm}0.47$, ME REG $4.31{\pm}1.58$ and SP REG $8.11{\pm}0.57$ fold compared to CON. And myogenin mRNA expression increased in MP REG $4.11{\pm}0.42$, ME REG $4.12{\pm}0.45$ and SP REG $6.50{\pm}0.61$ fold compared to CON. In conclusion, during the resistance exercise, providing mulberry and silkworm gives positive effect on AR mRNA and MRFs expression increase.

Molecular Phylogeny of the Family Tephritidae (Insecta: Diptera): New Insight from Combined Analysis of the Mitochondrial 12S, 16S, and COII Genes

  • Han, Ho-Yeon;Ro, Kyung-Eui
    • Molecules and Cells
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    • v.27 no.1
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    • pp.55-66
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    • 2009
  • The phylogeny of the family Tephritidae (Diptera: Tephritidae) was reconstructed from mitochondrial 12S, 16S, and COII gene fragments using 87 species, including 79 tephritid and 8 outgroup species. Minimum evolution and Bayesian trees suggested the following phylogenetic relationships: (1) A sister group relationship between Ortalotrypeta and Tachinisca, and their basal phylogenetic position within Tephritidae; (2) a sister group relationship between the tribe Acanthonevrini and Phytalmiini; (3) monophyly of Plioreocepta, Taomyia and an undescribed new genus, and their sister group relationship with the subfamily Tephritinae; (4) a possible sister group relationship of Cephalophysa and Adramini; and (5) reconfirmation of monophyly for Trypetini, Carpomyini, Tephritinae, and Dacinae. The combination of 12S, 16S, and COII data enabled resolution of phylogenetic relationships among the higher taxa of Tephritidae.