• Title/Summary/Keyword: r-DNA

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A Molecular Phylogenetic Study on Korean Alexandrium catenella and A. tamarense Isolates (Dinophyceae) Based on the Partial LSD rDNA Sequence Data

  • Kim, Keun-Yong;Kim, Chang-Hoon
    • Journal of the korean society of oceanography
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    • v.39 no.3
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    • pp.163-171
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    • 2004
  • Sequences of the large subunit ribosomal (LSD) rDNA D1-D2 region of Alexandrium catenella(=A. sp. cf. catenella) and A. tamarense isolates, which were collected along the Korea coasts, were analyzed to understand their phylogenetic relationships and geographical distributions. All A. catenella and A. tamarense isolates belonged to the A. tamarense/catenella/fundyense complex and were grouped with the North American and temperate Asian ribotypes, respectively, regardless of the presence or absence of a ventral pore in the first apical plate. A consistent and peculiar characteristic that differentiated the Alexandrium isolates was amplification of a second PCR product with a lower molecular weight in addition to the predicted one; ten A. catenella isolates belonging to the temperate Asian ribotype yielded this additional PCR product. Sequence alignment revealed that the shorter PCR product resulted from an unusual large deletion of 87 bp in the LSD rDNA D1 domain. The North American and temperate Asian ribotypes were prevalent along the Korean coasts without geographical separation. Given the high genetic homogeneity among widely distributed Alexandrium populations, each ribotype appeared to be pandemic rather than to constitute a distinct regional population.

Analysis of Endospore-forming Bacteria or Nitrogen-fixing Bacteria Community Isolated from Plants Rhizosphere in Dokdo Island (독도서식 식물근권에서 분리한 포자형성세균과 질소고정세균의 군집구조 분석)

  • Jeon, Seon-Ae;Sung, Hye-Ri;Park, Yu-Mi;Pak, Jae-Hong;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.37 no.3
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    • pp.189-196
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    • 2009
  • Bacteria were isolated from roots of plants belonging to family Solanaceae and Gramineae, inhabited in Dokdo island. Fifty six endospore-forming bacteria grown on tryptic soy broth (TSB) agar medium and 23 nitrogen-fixing bacteria (NFB) grown on nitrogen free agar medium were isolated, respectively. The isolates were partially identified by analyzing the 16S rDNA and categorized into phylogenetic groups. The 16S rDNA sequences of each identified isolates were compared with sequences of each type strains to analyze phylogenetic relationship by phylogenetic tree. As a result, endospore-forming bacteria and nitrogen-fixing bacteria were classified into 4 and 6 lineage groups, respectively. Among these isolated, 18 were presumed to be novel species candidates based on the similarity (lower than 98%) analysis of the l6S rDNA sequences.

Molecular Phylogeny of the Genera Staurastrum and Staurodesmus (Zygnematophyceae, Streptophyta) Based on Nuclear (18S rDNA) and Chloroplast Gene (atpB) Sequences (핵(18S rDNA)과 엽록체 유전자(atpB) 분석을 통한 Staurastrum속과 Staurodesmus속 (Zygnematophyceae, Streptophyta)의 분자 계통학적 연구)

  • Moon, Byeong-Ryeol;Lee, Ok-Min
    • ALGAE
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    • v.22 no.1
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    • pp.1-10
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    • 2007
  • To gain insights into the phylogenetic relationships of genus Staurastrum and Staurodesmus, we analyzed nuclearencoded small subunit rDNA of 82 strains, and chloroplast atpB gene sequences of 44 strains belonging to three genera (Staurastrum, Staurodesmus, Cosmarium). Excluding the Staurastrum muticum and S. orbiculare, forty five strains of genus Staurastrum formed a well supported clade. It was shown that with no cell wall sculpture and processes, these two species have a strong phylogenetic relationship with genus Staurodesmus. Therefore, it is strongly recommended to transfer Staurastrum without processes and cell wall sculpture into Staurodesmus. S. obsoletus is a taxa that is transferred from Cosmarium. But, from this study, it has shown a phylogenetic relationship with Cosmarium. Therefore, this species is strongly recommended to transfer back to Cosmarium instead of Staurodesmus. As it was studied before, genus Staurastrum has shown monophyletic. Since the genus taurodesmus groups with Cosmarium, they were shown to be polyphyletic.

Sequence Similarity of Nuclear 18S rDNA from Morphologically Different Blades of the Seaweed Porphyra pseudolinearis (Rhodophyta) (긴잎돌김 Porphyra pseudolinearis의 엽체형간 18S rDNA 염기서열 상동성)

  • Jin Long-Guo;KIM Young-Dae;KIM Myung-Sook;JIN Hyung-Joo;CHO Ji-Young;CHOI Jae-Suk;HONG Yong-Ki;KIM Hyung Geun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.6
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    • pp.496-500
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    • 2000
  • Partial fragments of nuclear 185 rDNAs from morphologically wide and narrow thalli of the seaweed Porphyra pseudolineazis were amplified and sequenced to compare their DNA homology. Both sequences of 311 base pairs showed $100{\%}$ identical each other. They showed $97.7{\%}$ similarity with a wild strain collected at Sodol in Kangwondo, and $99.4{\%}$ similarity with the GenBank accession number AB013185 of the Japanese P. pseudolinearis. Thus the morphological difference of wide and narrow blades might not be a classification criterion for the sub-species level of P. pseudolinearis.

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Comparison of Terminal-restriction Fragment Length Polymorphism (T-RFLP) Analysis and Sequencing of 16S rDNA Clones in marine sediments

  • Lee Jung-Hyun
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.15-21
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    • 2002
  • Terminal-restriction fragment length polymorphism (T-RFLP) analysis has been optimized by using in vitro model community composed of genomic DNAs of known bacterial strains and has been applied to assess the bacterial community structure in marine sediments. The specific fluorescence-labeled terminal restriction fragments (T-RFs) between 39 and 839 base long specifying each strain were precisely measured for known bacterial strains. The addition of a co-solvent (dimethylsulfoxide or glycerol) into PCR reactions has reduced differential PCR amplification. Comparative bacterial community structure was investigated for pristine and polluted sediments. A complex T-RFLP pattern showing complex bacterial community structure was obtained in the pristine sediment, whereas simple T-RFLP pattern (low bacterial diversity) was shown in polluted sediments where caged aquaculture has been conducted for several years. The results of T-RFLP analysis were compared with that of cloning and sequencing 16S rDNA clones from the same sediments. Sequence analysis of 16S rDNA clones (72) of the pristine sediment revealed a diverse collection of lineages, largely of the class Proteobacteria ($6\%$ alpha subdivision, $46\%$ gamma subdivision, $13\%$ delta subdivision, and $3\%$ epsilon subdivision), Nitrospina $(8\%)$, high G+C gram positive $(8\%)$, Verrucomicrobia $(7\%)$, and Planctomycetes $(6\%)$. In the contaminated sediments, 17 $(59\%)$ of the 16S rDNA clones (29) were related to Campylobacter and symbiont of Rimicaris exoculata belonging to epsilon subdivision of Proteobacteria. The results obtained indicated that T-RFLP analysis is a rapid and precise technique for comparative bacterial community analysis.

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Phylogenetic rind Taxonomic Status of the Phytoplasmas Associated with Water Dropwort (Oenanthe javanica DC) Disease in Korea and Japan

  • Jung, Hee-Young;Woo, Tae-Ha;Hibi, Tadaaki;Namba, Shigetou;Lee, Joon-Tak
    • The Plant Pathology Journal
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    • v.18 no.3
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    • pp.109-114
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    • 2002
  • To evaluate the phylogenetic and taxonomic status of the phytoplasmas associated with water dropwort (Oenanthe javanica DC) disease in Korea and Japan, their 16S rDNA was analyzed. DNAs extracted from water dropworts collected in Korea (Kyongnam province) and Japan (Chiba prefecture) affected by witches' broom and yellows were subjected to PCR using phytoplasma-specific primers, which amplified a 1.4-kbp fragment that included the 16S rDNA. Phytoplasmas were characterized by RFLP analysis using AluI, HaeIII, HhaI, KpnI, MseI, and RsaI restriction enzymes and by sequence analysis of the PCR products. The mater dropwort witches'broom (WDWB) and water dropwort yellows (WDY) 16S rDNA sequences were identical and closely related to onion yellows (OY, 99.9% identity), which belong to the aster yellows (AY) 16S-subgroup. However, the KpnI RFLP analyses clearly distinguished the WDY and WDWB phytoplasmas from the OY phytoplasma. The phylogenetic analysis based on 16S rDNA showed that WDWE and WDY phytoplasmas are members of a relatively homogeneous group that evolved from a common ancestor.

PCR Diagnosis of Entamoeba histolytica Cysts in Stool Samples

  • Moon, Joung-Ho;Cho, Shin-Hyeong;Yu, Jae-Ran;Lee, Won-Ja;Cheun, Hyeng-Il
    • Parasites, Hosts and Diseases
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    • v.49 no.3
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    • pp.281-284
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    • 2011
  • Amebiasis is a protozoan disease caused by Entamoeba histolytica and a potential health threat in areas where sanitation and hygiene are inappropriate. Highly sensitive PCR methods for detection of E. histolytica in clinical and environmental samples are extremely useful to control amebiasis and to promote public health. The present study compared several primer sets for small subunit (SSU) rDNA and histone genes of E. histolytica cysts. A 246 bp of the SSU rDNA gene of pure cysts contained in phosphate-buffered saline (PBS) and in stool samples was successfully amplified by nested PCR, using the 1,147-246 bp primer set, of the primary PCR products which were pre-amplified using the 1,147 bp primer as the template. The detection limit of the nested PCR using the 1,147-246 primer set was 10 cysts in both groups (PBS and stool samples). The PCR to detect histone gene showed negative results. We propose that the nested PCR technique to detect SSU rDNA can be used as a highly sensitive genetic method to detect E. histolytica cysts in stool samples.

Report on the Effects Lipemic Specimen in Anti-ds DNA Antibody Test (Anti-ds DNA 항체 검사 시 Lipemic 검체의 영향에 관한 보고)

  • Cheon, Jun Hong;Kim, Whe Jung;Kim, Sung Ho;Moon, Hyoung Ho;Yoo, Seon Hee
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.153-157
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    • 2014
  • Purpose: SLE (systemic lupus erythematosus) is an inflammatory autoimmune disease, characterized by various autoantibody. The detection of Anti double-stranded DNA (Anti-ds DNA) is important in the diagnostics of SLE, and include the American College of Rheumatology (ACR) diagnostic criteria for SLE. Also SLE disease activity and correlativity with the level Anti-ds DNA antibody have been reported and Anti-ds DNA antibody quantitative test is very useful for tracing before and after SLE treatment. When These Anti-ds DNA antibody test (Farr assay: $^{125}I$ labeled ds-DNA and bound Anti-ds DNA antibodies complex in serum is precipitated by ammonium sulfate and used to centrifugation, measured it) inhaled supernatant after centrifugation, a lipemic specimen does not facilitate the formation of precipitate and also occurs situation was inhaled with precipitate. To solve these problems, The Influence of the degree of lipemic specimen was evaluated. Materials and Methods: September 2012 to February 2013, We selected lipemic samples (n=81) of specimen commissioned by Anti-ds DNA antibody test. Lipemic samples were done pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) used a micro-centrifuge (Eppendorf Model 5415D). At the same time lipemic specimen and pre-treatment samples were performed Anti-ds DNA antibody test (Anti-ds DNA kit, Trinity Biotech, Ireland). Statistical analysis were analyzed Pearson's correlation coefficients and regression and paired t-test, and Difference (%). Results: Experimental group 1 (Lipemic Specimen Anti-ds DNA Ab concentration ${\leq}7IU/mL$) at y=0.368X+4.732, $R^2=0.023$, Pearson's correlation coefficient was 0.154, paired t-test (P=0.003), Difference (%) mean 65.7 and showed a statistically significant difference. Experimental group 2 (Lipemic Specimen Anti-ds DNA Ab concentration ${\geq}8IU/mL$) at y=0.983X+0.298, $R^2=0.994$, Pearson's correlation coefficient showed 0.997, paired t-test (P=0.181), Difference (%) mean -5.53 made no statistically significant difference. Conclusion: Lipemic sample of low Anti-ds DNA Ab concentration (2.5-7 IU/mL) and the result is obtained pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) were made a significant difference statistically. Anti-ds DNA is one of the primary auto-antibodies present in patients with SLE, and remain an important diagnostic test for SLE. Therefore, we recommend preprocessing (high-speed centrifugation: 14,000 rpm 5 mins) in order to exclude the influence of lipemic specimen.

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Phylogenetic Analysis of Dendropanax morbifera Using Nuclear Ribosomal DNA Internal Transcribed Spacer (ITS) Region Sequences (Internal transcribed spacer (ITS) region의 염기서열 분석에 의한 보길도산 황칠나무의 분자 계통학적 연구)

  • Shin, Yong Kook
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1341-1344
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    • 2016
  • Dendropanax morbifera is an endemic tree species of Korea, it is restricted to the southern parts of Korea. The internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) for Dendropanax morbifera grown at Bogil-do, Korea was determined. We investigated the sequence-based phylogenetic relationships of plants related and clarified its taxonomical position. The determined sequences consisted of 689 residues. ITS1 was 222 bp long while ITS2 was 233 bp long. The 5.8S rDNA was 160 bp long. The ITS region sequences of the Dendropanax species included in this study were obtained from GenBank. Oreopanax polycephalus was used as the outgroup. A pairwise alignment was calculated using the Clustal X program. A phylogenetic tree was constructed by the neighbor-joining method using the Tree view program. Sequence similarities among species including D. morbifera Bogil-do isolate showed the range 92.6 to 99.7% in sequence-based phylogenetic analysis using total 615 base pairs of ITS1, 5.8S rDNA and ITS2. D. morbifera Bogil-do isolate exhibited the highest degree of relatedness to D. chevalieri, sharing 99.7% ITS region similarity. D. morbifera Bogil-do isolate also showed to D. trifidus, sharing 99.4% ITS region similarity.

Binding Modes of New Bis-Ru(II) Complexes to DNA: Effect of the Length of the Linker

  • Kwon, Byung-Hyang;Choi, Byung-Hoon;Lee, Hyun-Mee;Jang, Yoon-Jung;Lee, Jae-Cheol;Kim, Seog-K.
    • Bulletin of the Korean Chemical Society
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    • v.31 no.6
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    • pp.1615-1620
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    • 2010
  • Bis-[dipyrido[3,2-$\alpha$:2',3'-c]phenazine)$_2$(1,10-phenanthroline)$_2Ru_2$]$^{2+}$ complexes (bis-Ru(II) complexes) tethered by linkers of various lengths were synthesized and their binding properties to DNA investigated by normal absorption and linear dichroism spectra, and fluorescence techniques in this study. Upon binding to DNA, the bis-Ru(II) complex with the longest linker (1,3-bis-(4-pyridyl)-propane), exhibited a negative $LD^r$ signal whose intensity was as large as that in the DNA absorption region, followed by a complicate $LD^r$ signal in the metal-to-ligand charge transfer region. The luminescence intensity of this bis-Ru(II) complex was enhanced. The observed $LD^r$ and luminescence results resembled that of the [Ru(1,10-phenanthroline)$_2$ dipyrido[3,2-$\alpha$:2',3'-c]phenazine]$^{2+}$ complex, whose dipyrido[3,2-$\alpha$:2',3'-c]phenazine (dppz) ligand has been known to intercalate between DNA bases. Hence, it is conclusive that both dppz ligands of the bis-Ru(II) complex intercalate. The binding stoichiometry, however, was a single intercalated dppz per ~ 2.3 bases, which violates the "nearest binding site exclusion" model for intercalation. The length between the two Ru(II) complexes may be barely long enough to accommodate one DNA base between the two dppz ligands, but not for two DNA bases. When the linker was shorter (4,4'-bipyridine or 1,2-bis-(4-pyridyl)-ethane), the magnitude of the LD in the dppz absorption region, as well as the luminescence intensity of both bis-Ru(II) complexes, was half that of the bis-Ru(II) complex bearing a long linker. This observation can be elucidated by a model whereby one of the dppz ligands intercalates while the other is exposed to the aqueous environment.