• Title/Summary/Keyword: small subunit ribosomal DNA

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Bacterial Diversity in the Mud Flat of Sunchon Bay, Chunnam Provice, by 16S rRNA Gene Analysis (16S rRNA 유전자 분석에 의한 전남 순천만 갯벌의 세균 다양성)

  • 이명숙;홍순규;이동훈;배경숙
    • Korean Journal of Microbiology
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    • v.37 no.2
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    • pp.137-144
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    • 2001
  • In order to investigate the diversity of bacterial community in the mud flat of Sunchon Bay, Chunnam province, diversity of amplified 16S rDNA was examined. Total DNA was extracted from sediment soils and 16S rDNAs were amplified using PCR primers based on the universally conserved sequences in bacteria. Clonal libraries were constructed and 111 clones were examined by amplified rDNA restriction analysis (ARDRA) using HaeIII. Clones were clustered based on restriction patterns using computer program, GelCompar II. One hundred different RFLP types were detected from 111 clones. The 20 clones were selected and sequenced according to dendrograms derived from ARDRA, to cover most of the bacterial diversity in the clone libraries. None of the clones were identical to any representatives in the Ribosomal Database Project small subunit RNA databases and GenBank. All sequences showed between 77 and 96.8% similarity to the known 16s rRNA sequence from cultured organisms. The 20 clones sequenced fell into seven major lineages of the domain Bacteria: alpha-, delta-, gamma-Proteobacteria, low G+C Gram positive bacteria, high G+C Gram positive bacteria, Sphingobacteria (Cytophaga) and Cyanobacteria (chloroplast). Among the clones, the Proteobacteria were dominant.

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Isolation and Identification of Acanthamoeba in a Contact Lens Storage Case

  • Moon, Eun-Kyung
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.324-327
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    • 2012
  • Acanthamoeba is a free-living amoeba that causes human infections, and recently the incidence of amoebic keratitis has increased among contact lens wearers. In order to investigate Acanthamoeba contamination of contact lens storage cases, a short survey was performed on 57 contact lens wearers, and Acanthamoeba was found in one contact lens storage case. To diagnose Acanthamoeba, the 18s small subunit ribosomal DNA (18s rDNA) gene was amplified by polymerase chain reaction (PCR), and subsequently, the isolate was identified as A. lugdunensis. This species was originally isolated from a freshwater pool in France, and was reported recently to be a cause of amoebic keratitis. This observation indicates the need for a large survey to investigate the extent of Acanthamoeba contamination, and suggests that contact lens wearers be aware of the importance of hygiene and of the implications of Acanthamoeba infection.

Study on the variation of cellular physiology of Escherichia coli during high cell density cultivation using 2-dimensional gel electrophoresis

  • Yun, Sang-Seon;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.219-222
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    • 2000
  • Physiological changes of Escherichia coli during the fed-batch fermentation process were characterized in this study. Overall cellular protein samples prepared at the different stage of fermentation were separated by 2-dimensional gel electrophoresis (2-DE), and differently expressed 15 proteins, Phosphotransferase enzyme I, GroEL, Trigger factor, ${\beta}$ subunit of ATP synthase, Transcriptional regulator KDGR, Phosphoglycerate mutase 1, Inorganic pyrophosphatase, Serine Hydroxymethyl-transferase, ${\alpha}$ subunit of RNA polymerase, Elongation factor Tu, Elongation factor Ts, Tyrosine-tRNA ligase, DnaK suppressor protein, Transcriptional elongation factor, 30S ribosomal protein S6 were identified using matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). When bacterial cells grow to high cell density, and IPTG-inducible heterologous protein is produced, expression level of overall cellular proteins was decreased. According to their functions in the cell, identified proteins were classified into three groups, proteins involved in transport process, small-molecule metabolism, and synthesis and modification of macromolecules.

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Penicillium vietnamense sp. nov., the First Novel Marine Fungi Species Described from Vietnam with a Unique Conidiophore Structure and Molecular Phylogeny of Penicillium Section Charlesia

  • Nguyen, Van Duy;Pham, Thu Thuy
    • Mycobiology
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    • v.50 no.3
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    • pp.155-165
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    • 2022
  • Penicillium vietnamense sp. nov. was isolated from Nha Trang Bay, Vietnam in June 2017. It is phylogenetically distinct from the sister species of Penicillium section Charlesia series Indica based on multi-locus sequence typing results using internal transcribed spacer, large subunit ribosomal RNA, b-tubulin, calmodulin, and RNA polymerase II second largest subunit regions. It showed strong growth on Czapek yeast autolysate agar at 37 ℃, a strong acid production on Creatine sucrose agar, and produced short stipes, small vesicles, and subglobose to globose conidia delicately roughened with very short ridges. As the first novel marine fungi species described from Vietnam and discovered in a unique environment, the data could be significant for understanding the taxonomy and geographical distribution of marine fungi in tropical coastal systems such as Vietnam.

PCR and RFLP variation of conserved region of small subunit ribosomal DNA among Acanthamoeba isolates assigned to either A. castellanii or A. polyphaga (카스텔라니가시아메바 혹은 대식가시아메바로 분류된 분리주간의 ribosomaIDNA conserved region의 PCR-RFLP의 다양성)

  • 공현희;정동일
    • Parasites, Hosts and Diseases
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    • v.34 no.2
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    • pp.127-134
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    • 1996
  • Twelve isolates of Accnthamoebc app. assigned to either A. castellanii or A. poIMphoSa, and type strains of A. culbefsoni, A. henIWi, A. pqkestinefiE, and A. astronyxi,s were examined by restriction fragment length polymorphism (RFLP) of a conserved region of small subunit ribosomal RMh gene (ssu rDNA) amplified by polymerase chain reaction (PCR). The PCR products of the isolates measured approximately 910-930 bp, except for that of A. astronyxis which was extraordinarily long, approximately 1,170 bp. Average of estimated sequence divergence of the amplified DNA among the isolates assigned to A. castellanii was 9.8% whereas that among the isolates assigned to A. polvphusn 9.6%. The maximum intraspecific sequence divergence among the isolates assigned to A. costellanii was observed between the Chang and Ma strains (17.3%) while that among the isolates assigned to A. poIWphosa was observed between KA/S3 and KA/S7 strains (16.1%). The both maximum sequence divergences were much greater than the minimum interspecific sequence divergence between A. cnstellnnii and A. polwphasa (2.6%) which appeared between the Castellani (or CCAP 1501/12 g) and KA/S3 strains. The PCR-RFLP patterns of A. culbertsoni, A. healyi, A. palestinensis, and A. ostronvxis were quite diverse from one another and from those of isolates assigned to either A. castellanii or A. polyphoga. It is suggested that taxonomic validity of the isolates assigned to either A. castellnnii or A. polyphoga should be reevaluated.

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Molecular Characterization of Small-Spored Alternaria Species (소형의 포자를 형성하는 Alternaria 균류의 분자생물학적 특징)

  • Kim, Byung-Ryun;Park, Myung-Soo;Cho, Hye-Sun;Yu, Seung-Hun
    • Research in Plant Disease
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    • v.11 no.1
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    • pp.56-65
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    • 2005
  • To establish taxonomic system of morphologically similar species of small-spored Alternaria, phylogenetic analysis of internal transcribed spacer (ITS 1, ITS 2 and 5.8S rDNA) and mitochondrial small subunit (mt SSU) rDNA sequences and URP-PCR fingerprinting analysis from 11 species ofAlternaria were performed. Phylogenetic analysis of ITS and mt SSU rDNA sequences revealed that 10 out of 11 species of the smallspored Alternaria were phylogenetically identical with a bootstrap value of 100%. A. infectoria only was phylogenetically differentiated from the other species. The results suggest that the 10 small-spored Alternaria species are very closely related evolutionally and the markers can not be used for differentiation of the smallspored Alternaria species. URP-PCR fingerprinting analysis from eleven species of smallspored Alternaria using 10 URP primers showed that it was possible to differentiate the species, although genetic similarities were found among the species. The Alternaria sp. from common pokeweed could be distinguished from other species by URP-PCR analysis, and it was considered as a new species. A. infectoria could be easily distinguished from the other 10 species by phylogenetic analysis of ITS and mt SSU rDNA sequences and the URPPCR fingerprinting analysis.

A riboprinting scheme for identification of unknown Acanthamoeba isolates at species level

  • Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.40 no.1
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    • pp.25-31
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    • 2002
  • We describe a riboprinting scheme for identification of unknown Acanthamoeba isolates at the species level. It involved the use of PCR-RFLP of small subunit ribosomal RNA gene (riboprint) of 24 reference strains by 4 kinds of restriction enzymes. Seven strains in morphological group I and III were identified at species level with their unique sizes of PCR product and riboprint type by Rsa 1. Unique RFCP of 17 strains in group II by Dde I. Taq I and Hae III were classified into: (1) four taxa that were identifiable at the species level. (2) a subgroup of 4 taxa and a pair of 2 taxi that were identical with each other. and (3) a species complex of 7 taxa assigned to A. castellanii complex that were closely related. These results were consistent with those obtained by 18s rDNA sequence analysis. This approach provides an alternative to the rDNA sequencing for rapid identification of a new clinical isolate or a large number of environmental isolates of Acanthamoeba.

Identification of Arbuscular Mycorrhizal Fungi from Botrychium ternatum Native in Korea

  • Lee, Jun-Ki;Eom, Ahn-Heum;Lee, Sang-Sun
    • Mycobiology
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    • v.32 no.4
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    • pp.179-185
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    • 2004
  • Arbuscular mycorrhizal fungi were observed in Botrychium ternatum native in Korea. The partial small subunit(SSU) of ribosomal DNA gene from the fern roots was amplified with primers AM1/NS31. Nucleotides sequence analysis of the clones revealed that two fragments were close to Glomus proliferum and G. sinuosum. The other three DNA fragments were close to those of G. proliferum with the relatively low similarities($92{\sim}95%$) and speculated to be originated from three different species of Glomus(GLA006, GLA016, and GLA032). Five different nucleotide sequences close to three AM fungal species were found in the roots of B. ternatum native in Korea.

Blooms of the woloszynskioid dinoflagellate Tovellia diexiensis sp. nov. (Dinophyceae) in Baishihai Lake at the eastern edge of Tibetan Plateau

  • Zhang, Qi;Zhu, Huan;Hu, Zhengyu;Liu, Guoxiang
    • ALGAE
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    • v.31 no.3
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    • pp.205-217
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    • 2016
  • Freshwater red tides due to dinoflagellates have caused spectacular and regular “summer reddening” in recent years in Baishihai Lake, a temperate, meromictic, meso- or oligotrophic, high-altitude, landslide-dammed, deep lake located at the eastern edge of Tibetan Plateau in China. Based on morphological and molecular analyses, the causative organism has been identified as a new woloszynskioid dinoflagellate, Tovellia diexiensis Q. Zhang et G. X. Liu sp. nov. The vegetative cells are 20-32 μm long and 16-24 μm wide. They have a hemispherical episome and a broadly rounded hyposome with a short characteristic antapical spine. Usually cells are bright red due to the presence of numerous red-pigmented bodies, which often masked the yellowish green discoid chloroplasts. The amphiesma of motile cells comprise mainly quadrilateral, pentagonal or hexagonal thin plates, arranged in 4-5 latitudinal series on the episome, 1 in the cingulum and 4 on the hyposome. Molecular phylogenies based on small subunit ribosomal DNA and large subunit ribosomal DNA (LSU) indicate T. diexiensis from Baishihai Lake to belong to the family Tovelliaceae, which was monophyletic in our LSU phylogenies. During the bloom-forming period in 2005, cell density of T. diexiensis reached 9.15 × 105 cells L−1. Astaxanthin and its diester were found to be the major pigments in T. diexiensis, resulting in a characteristic blood-red color of the water in Baishihai Lake.

Molecular Phylogenetic Relationships Within the Genus Alexandrium(Dinophyceae) Based on the Nuclear-Encoded SSU and LSU rDNA D1-D2 Sequences

  • Kim, Choong-Jae;Sako Yoshihiko;Uchida Aritsune;Kim, Chang-Hoon
    • Journal of the korean society of oceanography
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    • v.39 no.3
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    • pp.172-185
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    • 2004
  • LSU rDNA D1-D2 and SSU rDNA genes of 23 strains in seven Alexandrium (Halim) species, A. tamarense (Lebour) Balech, A. catenella (Whedon et Kofoid), A. fraterculus (Balech) Balech, A. affine (Inoue et Fukuyo) Balech, A. insuetum Balech, A. pseudogonyaulax (Biecheler) Horiguchi ex Yuki et Fukuyo and A. tamiyavanichii Balech, were sequenced and the data were used for molecular phylogenetic analysis. The sequence data revealed 11 and 7 ribotypes in the LSU rDNA D1-D2 region and 4 and 17 ribotypes in the SSU rDNA region of A. catenella and A. tamarense, respectively. Other Alexandrium species had also 1 to 5 ribotypes in the two regions. With the exception of CMC2 and CMC3 of A. catenella, all A. tamarense and A. catenella strains had a common ribotype, a functionally expressed rRNA gene (here termed type A), in both gene regions. In addition to the functionally expressed gene, several pseudogenes were obtained that were found to be good tools to analyze the population designation of regional isolates by grouping them according to shared ribotypes. From the phylogenetic analysis of the sequence data determined in this study and retrieved from GenBank, the genus Alexandrium was divided into 14 groups: 1) A. tamarense, 2) A. excavatum, 3) A. catenella, 4) Tasmanian A. tamarense, 5) A. affine (and/or A. concavum), 6) Thai A. tamarense, 7) A. tamiyavanichii, 8) A. fraterculus, 9) A. margalefii, 10) A. andersonii, 11) A. ostenfeldii, 12) A. minutum (or A. lusitanicum), 13) A. insuetum, and 14) A. pseudogonyaulax. The SSU rDNA gene sequence of A. fundyense was so similar to those of A. tamarense used in this study that the two species were difficult to discriminate each other. A. tamiyavanichii was closest to the A. tamarense strain isolated in Thailand and close to the long chain-forming species of A. affine and A. fraterculus. The phylogenetic tree showed that A. margalefii, A. andersonii, A. ostenfeldii, A. minutum and A. insuetum constituted the basal relative complex, and that A. pseudogonyaulax is an ancestral taxon in the genus Alexandrium.