• Title/Summary/Keyword: ITS rDNA

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Seasonal and Spatial Diversity of Picocyanobacteria Community in the Great Mazurian Lakes Derived from DGGE Analyses of 16S rDNA and cpcBA-IGS Markers

  • Jasser, Iwona;Krolicka, Adriana;Jakubiec, Katarzyna;Chrost, Ryszard J.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.739-749
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    • 2013
  • The seasonal and spatial diversity of picocyanobacteria (Pcy) in lakes of the Great Mazurian Lakes (GLM) system was examined by DGGE analysis of molecular markers derived from the 16S-23S internal transcribed spacer (ITS) of the ribosomal operon and the phycocyanin operon (cpcBA-IGS). The study of nine lakes, ranging from mesotrophy to hypereutrophy, demonstrated seasonal variance of Pcy. The richness and Shannon diversity index calculated on the basis of both markers were higher in spring and lower in early and late summer. No statistically significant relationships were found between the markers and trophic status of the studied lakes or Pcy abundance. There were, however, statistically significant relationships between the diversity indices and sampling time. The analysis pointed to a different distribution of the two markers. The ITS marker exhibited more unique sequences in time and space, whereas a greater role for common and ubiquitous sequences was indicated by the cpcBA-IGS data. Examination of the Pcy community structure demonstrated that communities were grouped in highly similar clusters according to sampling season/time rather than to the trophic status of the lake. Our results suggest that time is more important than trophic status in shaping the diversity and structure of Pcy communities. The seasonal changes in picocyanobacteria and differences in diversity and community structures are discussed in the context of well-established ecological hypotheses: the PEG model, intermediate disturbance hypothesis (IDH), and horizontal gene transfer (HGT).

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 Molecular Characterization of a New CRT Binding Factor Gene from Capsella bursa-pastoris

  • Wang, Xinglong;Liu, Li;Liu, Sixiu;Sun, Xiaoqing;Deng, Zhongxiang;Pi, Yan;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.37 no.5
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    • pp.538-545
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    • 2004
  • A new CRT binding factor (CBF) gene designated Cbcbf25 was cloned from Capsella bursa-pastoris, a wild grass, by the rapid amplification of cDNA ends (RACE). The full-length cDNA of Cbcbf25 was 898 bp with a 669 bp open reading frame (ORF) encoding a putative DRE/CRT (LTRE)-binding protein of 223 amino acids. The predicted CbCBF25 protein contained a potential nuclear localization signal (NLS) in its N-terminal region followed by an AP2 DNA-binding motif and a possible acidic activation domain in the C-terminal region. Bioinformatic analysis revealed that Cbcbf25 has a high level of similarity with other CBF genes like cbf1, cbf2, and cbf3 from Arabidopsis thaliana, and Bncbf5, Bncbf7, Bncbf16, and Bncbf17 from Brassica napus. A cold acclimation assay showed that Cbcbf25 was expressed immediately after cold triggering, but this expression was transient, suggesting that it concerns cold acclimation. Our study implies that Cbcbf25 is an analogue of other CBF genes and may participate in cold-response, by for example, controlling the expression of cold-regulated genes or increasing the freezing tolerance of plants.

An Ecological Analysis of Lichens Distributed in Rocks of Coast and Field in U-do by Molecular Technique (분자생물학적 기법에 의한 우도해안과 노지암석에 분포하는 지의류의 생태학적 분석)

  • Gang, Hyeong-Il;Yun, Byeong-Jun;Kim, Seong-Hyeon;Sin, Deok-Ja;Kim, Hyeon-U;Heo, Jae-Seon;Gang, Ui-Seong;O, Gye-Heon;Go, Yeong-Jin
    • Microbiology and Biotechnology Letters
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    • v.32 no.4
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    • pp.334-340
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    • 2004
  • This study was initially performed to obtain fundamental data on lichen distribution depending on environmental condiLichens distributed on coastal rocks atleast 20 m far away from the coast were also investigated. Analyof morphological characteristics and rDNA ITS clones revealed that lichens containing 9 families and 15 genera were distributed in coastal rocks of V-do, while lichens containing 10 families and 14 genera were in field rocks, demonstrating little difference of diversity between both sites. Foliose lichens such as PhaeophysPhyscia, Pyxine belonging to the family Physciaceae were representatives distributed in coastal rocks of V-do, along with Xanthoparmelia belonging to Parmeliaceae, fruticose lichens such as Ramalina to Ramaliand crustose lichen Lecanora to Lecanoraceae. In contrast, foliose lichen Cladonia belonging to the family Cladoniaceae, fruitcose lichen Stereocaulon to Streocaulaceae, and crustose lichen Porpidia to Porpidwere found only on the surface of field rocks. Crustose lichens containing genera Caloplaca, CanDirinaria, Graphis, Rhizocarpon, and Pertusaria were rarely distributed in coastal rocks, and most of them were also found in field rocks.

Occurrence of Sclerotium Rot of Cucumber Caused by Sclerotium rolfsii (Sclerotium rolfsii에 의한 오이 흰비단병 발생)

  • Kwon, Jin-Hyeuk;Lee, Sang-Dae;Choi, Okryun;Shen, Shun-Shan;Shim, Hong-Sik
    • Research in Plant Disease
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    • v.19 no.3
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    • pp.229-232
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    • 2013
  • Sclerotium rot of cucumber (Cucumis sativus L.) occurred at the experimental field of Gyeongsangnam-do Agricultural Research and Extension Services in July 2012. The typical symptoms included wilt, rot, and water-soaking on stems and fruits and severely infected plants eventually died. White mycelial mats spread over lesions, and then sclerotia were formed on fruit and near soil line. The sclerotia were globoid in shape, white to brown in color and 1-3 mm in size and the hyphal width was 4-8 ${\mu}m$. The optimum temperature for mycelial growth and sclerotia formation on PDA was $30^{\circ}C$. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. For further identification, the complete internal transcribed spacer (ITS) rDNA region was amplified and sequenced. On the basis of mycological characteristics, ITS rDNA region comparison, and pathogenicity to host plants, this fungus was identified as Sclerotium rolfsii Saccardo. This is the first report of sclerotium rot on cucumber caused by S. rolfsii in Korea.

Synthesis and Characterization of a Hydroxylated Dendrimeric Gene Delivery Carrier

  • Kim, Tae-Il;Bai, Cheng-Zhe;Park, Jong-Sang
    • Bulletin of the Korean Chemical Society
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    • v.28 no.8
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    • pp.1317-1321
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    • 2007
  • Arginine conjugated PAMAM dendrimer, PAMAM-R was modified with propylene oxide via hydroxylation of primary amines of arginine residues. About 49 amines were detected to be converted to amino alcohols by 1H NMR. The newly synthesized polymer, PAMAM-R-PO was able to completely retard pDNA from a charge ratio of 2. The average diameter of PAMAM-R-PO polyplex was found to be 242 nm at a charge ratio of 30. The Zeta-potential value of PAMAM-R-PO polyplex was able to reach 20-30 mV over a charge ratio of 10. PAMAM-R-PO indicated higher cell viability than unmodified PAMAM-R on HeLa and 293 cells because of its hydroxylated amines. Transfection experiments on 293 cells showed that the transfection efficiency of PAMAM-R-PO was found to be 1.5-1.9 times higher than that of PEI25kDa at a charge ratio of 30. The polymer eventually displayed about 2 times greater transfection efficiency than PAMAM-R at the same charge ratio in the absence of serum. Therefore, we concluded that the modification of primary amines of PAMAMR to amino alcohols gives positive effects such as reduced cytotoxicity and enhanced transfection efficiency on 293 cells for gene delivery potency of PAMAM-R.

Genotype and Phenotype of Echinococcus granulosus Derived from Wild Sheep (Ovis orientalis) in Iran

  • Eslami, Ali;Meshgi, Behnam;Jalousian, Fatemeh;Rahmani, Shima;Salari, Mohammad Ali
    • Parasites, Hosts and Diseases
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    • v.54 no.1
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    • pp.55-60
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    • 2016
  • The aim of the present study is to determine the characteristics of genotype and phenotype of Echinococcus granulosus derived from wild sheep and to compare them with the strains of E. granulosus sensu stricto (sheep-dog) and E. granulosus camel strain (camel-dog) in Iran. In Khojir National Park, near Tehran, Iran, a fertile hydatid cyst was recently found in the liver of a dead wild sheep (Ovis orientalis). The number of protoscolices (n=6,000) proved enough for an experimental infection in a dog. The characteristics of large and small hooks of metacestode were statistically determined as the sensu stricto strain but not the camel strain (P=0.5). To determine E. granulosus genotype, 20 adult worms of this type were collected from the infected dog. The second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA (rDNA) and cytochrome c oxidase 1 subunit (COX1) of the mitochondrial DNA were amplified from individual adult worm by PCR. Subsequently, the PCR product was sequenced by Sanger method. The lengths of ITS2 and COX1 sequences were 378 and 857 bp, respectively, for all the sequenced samples. The amplified DNA sequences from both ribosomal and mitochondrial genes were highly similar (99% and 98%, respectively) to that of the ovine strain in the GenBank database. The results of the present study indicate that the morpho-molecular features and characteristics of E. granulosus in the Iranian wild sheep are the same as those of the sheep-dog E. granulosus sensu stricto strain.

Taxonomic status of Goodyera rosulacea (Orchidaceae): molecular evidence based on ITS and trnL sequences (로젯사철란(Goodyera rosulacea: Orchidaceae)의 분류학적 위치: ITS와 trnL 염기서열에 의한 분자적 증거)

  • Lee, Chang Shook;Eom, Sang Mi;Lee, Nam Sook
    • Korean Journal of Plant Taxonomy
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    • v.36 no.3
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    • pp.189-207
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    • 2006
  • Goodyera rosulacea, which is morphologically similar to G. repens, is described recently as a new species based on its distinct morphological characters such as rosette-formed leaves, short rhizome and habitat. To verify the taxonomic identity of G. rosulacea and its taxonomic relationship within Korean Goodyera taxa, sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA and the trnL region of cpDNA from 24 accessions including 1 outgroup accession were analyzed. Aligned sequences were analyzed using maximum parsimony and distance method, and the taxonomic identity and the taxonomic relationships among the related taxa were estimated by the existence of private marker gene and the phylogenetic tree of the aligned sequences. Molecular data indicate that G. rosulacea gas several private marker genes and shows monophyly in phylogenetic trees of both ITS and trnL sequences. the pairwise distance between G. rosulacea and the orher taxa of Korean Goodyera was 3.49-6.68% for ITS region and 5.05-9.53% for trnL region, indicating that G. rosulacea could be treated as an independent species. Therefore, our molecular data support the taxonomic of G. rosulacea as a distinct species of Korea. In phylogenetic trees, G. rosulacea formed same clade with G. repens, which has similar morphological characters with G. rosulacea, and showed the lowest pairwise distance with G. repens among Korean Goodyera taxa. These molecular data sugguested that G. rosulacea and G. repens are closely related taxa.

Detection of Biodegradative Genes in Oil Contaminated Soil Microbial Community by Oligonucleotide Microarray (Oligonucleotide Microarray를 이용한 유류 오염 토양 미생물 군집내 난분해성 화합물 분해 유전자의 검출)

  • Lee Jong-Kwang;Kim Hee;Lee Doo-Myoung;Lee Seok-Jae;Kim Moo-Hoon
    • Journal of Soil and Groundwater Environment
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    • v.11 no.1
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    • pp.1-6
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    • 2006
  • The analysis of functional population and its dynamics on the environment is essential for understanding bioremediation in environment. Here, we report a method for oligonucleotide microarray for the monitoring of aliphatic and aromatic degradative genes. This microarray contained 15 unique and group-specific probes which were based on 100 known genes involved pathways in biodegradation. Hybridization specificity tests with pure cultures, strain Pseudomonas aeruginosa KCTC 1636 indicated that the designed probes on the arrays appeared to be specific to their corresponding target genes. It was found that the presence of 8 genes encoding alkane, naphthalene, biphenyl, pyrene (PAH ring-hydroxylating) degradation pathway could be detected in oil contaminated soil sample. Therefore, the findings of this study strongly suggest that oligonucleotide microarray is an effective diagnostic tool for evaluating biodegradation capability in oil contaminated subsurface environment.

Purification and Fluorometric Analysis of Leucine-Responsive Regulatory Protein from Escherichia coli (대장균 Leucine-Responsive Regulatory Protein의 정제 및 형광분광학적 특성 분석)

  • Lee, Chan-Yong;Kim, Sung-Chul;Seo, Cho-Hee
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.104-108
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    • 2010
  • We describe the construction of derivatives of wild type and mutant lrp genes that encode 6XHis-tag Lrps. These derivatives of wild type and mutant Lrp could be useful for in vitro studies including Lrp conformational changes. We show that 6XHis-tag Lrp wild type and 6XHis-tag Lrp R145W bind with similar patterns in vitro to 21 bp duplex DNA containing the consensus sequences of Lrp sites of upstream of the ilvIH operon. In addition, we report here the 6XHis-tag Lrp R145W is useful to investigate the conformational changes of Lrp in solution by using its own intrinsic fluorescence characteristics.