• 제목/요약/키워드: 18S rDNA sequence analysis

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한국산 잇바디돌김 (Porphyra dentata)의 핵 18S rDNA 염기선열 분석 (Sequence Analysis of Nuclear 18s rDNA from Porphyra dentata (Rhodophyta) in Korea)

  • ;김명숙;조지영;진형주;홍용기
    • 생명과학회지
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    • 제12권4호
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    • pp.427-432
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    • 2002
  • 잇바디돌김(Porptyra dentata)을 대상으로 핵의 18S ribo-somal RNA를 지령하는 유전자 즉 18S rDNA 유전자를 증폭하고, 염기서열분석을 수행하였다. 전체 18S rDNA의 exon 영역 크기는 1822 bp, intron 영역의 크기는 512 bp였다. 이들 exon과 intron 영역의 G+C함량은 각각 49%와 55%씩 나타내었다. 일본산 잇바디돌김(CenBank accession number: AB013183)의 exon 영역과의 비교에서 상동성이 97.1%에 도달하였다. 568번과 569번 염기사이의 upstream에 위치하는 intron 영 역에서는 AB013183과 52.1%의 상동성을 보였다.

지황(地黃)의 18S rRNA 유전자 염기서열의 분석 및 분류학적 연구 (Determination of the DNA Sequence of the 18S rRNA Gene of the Rehmannia glutinosa and Its Phylogenetic Analysis)

  • 배은하;신동민;배영민
    • 대한본초학회지
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    • 제21권2호
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    • pp.9-13
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    • 2006
  • Objectives : To determine the DNA sequence of the 18S rRNA gene of the Rehmannia glutinosa and analyze it phylogenetically Methods : Dried root of the Rehmannia glutinosa was ground with a mortar and pestle. Glass beads(0.5 mm in diameter), TE buffer and SDS solution were added to that. The mixture was vortexed vigorously and extracted with the mixture of phenol, chloroform and isoamyl alcohol and with the mixture of the chloroform and isoamyl alcohol. The nucleic acids were precipitated with ethanol and resuspended in TE buffer. Contaminating RNA was digested with RNAse A and the DNA was purified further with the Geneclean Turbo Kit. This DNA was used as a template for amplification of the 18S rRNA gene by PCR. The PCR product was cloned in the pBluescript SK II plasmid by blunt-end ligation and the DNA sequence of the insert was determined. This DNA sequence was analyzed phylogenetically by the BLAST program. Results and Conclusion : Vortexing the ground powder of the dried plant root with glass beads during cell lysis improved recovery of DNA. The DNA sequence of the Rehmannia glutinosa 18S rRNA gene was determined and deposited at the GenBank as the accession number DQ469606. Phylogenetic analysis of that sequence showed the relationship between the members of the family of Scrophulariaceae and also the close relationship of the Buddleja davidii to the members of the Scrophulariaceae family.

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한국 주변해역 가리비로부터 분리한 18S rDNA의 염기서열 분석 (Sequence Analysis of the 18S rDNA from Scallops Collected around Korean Sea)

  • 김미정;;진형주;조지영;박중연;장영진;홍용기
    • 한국수산과학회지
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    • 제34권2호
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    • pp.137-144
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    • 2001
  • Sequences of partial 18S rDNA have been analyzed to elucidate genetic diversity of scallops collected around Korean sea, The scallops used in genetic comparison are Argopecten irradians concentricus, Amusium japonicum japonicum, Chlamys farreri farreri, Chlamys (Swiftopecten) swifti and Patinopecten yessoensis. The 18S rDNA sequences were aligned by Clustalx program. Phylogenetic tree was drawn by Treecon program, The scallops were divided into two groups-the Family Pectinidae containing A. japonicum japonicum and the Family Propeamussiidae containing Argopecten, Chlamys and Patinopecten genera. The Family Propeamussiidae was also divided into the Supergenera Aequipecten containing A. irradians concentricus and Supergenera Chlamys containing C. farreri farreri, C. swifti and P. yessoensis. The species of C. swifti was closer to the P. yessoensis rather than C. farreri farreri in respect to nuclear 18S rDNA sequence.

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벚나무 빗자루병균 Taphrina wiesneri의 유전적 특성 (Genotypic Characterization of Cherry Witches' Broom Pathogen Taphrina wiesneri Strains)

  • 서상태;정수지;이승규;김경희
    • 식물병연구
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    • 제17권1호
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    • pp.99-101
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    • 2011
  • 자낭균인 Taphrina wiesneri는 한국의 공원과 가로수에 주로 식재되는 왕벚나무에 빗자루병을 일으키는 병원균이다. 한국과 일본에서 분리한 13개의 병원균에 대해 18S rDNA 염기서열 분석을 통한 계통학적 분석과 rDNA-IGS 영역에 대한 RFLP 분석을 실시하였다. 18S rDNA 염기서열 분석을 통한 계통도 분석결과 병원균은 2그룹으로 분류되었다. Hha I 제한효소를 이용한 rDNA-IGS 영역에 대한 RFLP 분석결과 B, C, D, G 4개의 패턴으로 나타났으며, 그중 G 패턴은 새로운 패턴이었다.

한국산 방사무의김 (Porphyra yezoensis)의 핵 18S rDNA 염기서열 분석 (Sequence Analysis of Nuclear 18S rDNA from the Seaweed Porphyra yezoensis (Rhodophyta) in Korea)

  • ;김명숙;최재석;조지영;진형주;홍용기
    • 한국수산과학회지
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    • 제35권6호
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    • pp.633-638
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    • 2002
  • Nuclear 18S ribosomal RNA gene (185 rDNA) from the aquaculturable seaweed Porphya yezoensis (Bangiales, Rhodophyta) was amplified using the polymerase chain reaction and its sequence was analysed. Complete 185 rDNA has an 1823 bp exon and a 514 bp intron. The G+ C contents of exon and intron were $48\%$ and $51.4\%$, respectively. The exon sequence showed $99.5\%$ homology to the GenBank accession number AB013177 of the Japanese p. yezoensis. The intron region that was inserted upstream between 568 and 1083 showed $93.4\%$ homology to the AB013177.

한국산 참김 (Porphya tenera)의 핵 18S rDNA염기서열 분석 (Sequence Analysis of Nuclear 18S rDNA from Porphya tenera (Rhodophyta) in Korea)

  • ;김명숙;최재석;조지영;진덕희;홍용기
    • 한국수산과학회지
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    • 제36권1호
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    • pp.35-38
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    • 2003
  • Nuclear 18S ribosomal RNA gene (185 rDNA) from the aquaculturable seaweed Porphya tenera (Bangiales, Rhodophyta) was amplified using the polymerase chain reaction and its sequence was analysed. Complete 185 rDNA has an 1,822 bp exon and a 510 bp intron. The G+C contents of exon and intron were $48.68\%\;and\;54,90\%,$ respectively. The exon sequence showed $99.6\%$ homology to the GebBank accession number AB029880 of the Japanese P. tenera. The intron region that is inserted upstream between 568 and 1,079 showed $43.6\%$ homology to the AB029880.

Application of rDNA-PCR Amplification and DGGE Fingerprinting for Detection of Microbial Diversity in a Malaysian Crude Oil

  • Liew, Pauline Woan Ying;Jong, Bor Chyan
    • Journal of Microbiology and Biotechnology
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    • 제18권5호
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    • pp.815-820
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    • 2008
  • Two culture-independent methods, namely ribosomal DNA libraries and denaturing gradient gel electrophoresis (DGGE), were adopted to examine the microbial community of a Malaysian light crude oil. In this study, both 16S and 18S rDNAs were PCR-amplified from bulk DNA of crude oil samples, cloned, and sequenced. Analyses of restriction fragment length polymorphism (RFLP) and phylogenetics clustered the 16S and 18S rDNA sequences into seven and six groups, respectively. The ribosomal DNA sequences obtained showed sequence similarity between 90 to 100% to those available in the GenBank database. The closest relatives documented for the 16S rDNAs include member species of Thermoincola and Rhodopseudomonas, whereas the closest fungal relatives include Acremonium, Ceriporiopsis, Xeromyces, Lecythophora, and Candida. Others were affiliated to uncultured bacteria and uncultured ascomycete. The 16S rDNA library demonstrated predomination by a single uncultured bacterial type by >80% relative abundance. The predomination was confirmed by DGGE analysis.

Molecular Analysis of Complete SSU to LSU rDNA Sequence in the Harmful Dinoflagellate Alexandrium tamarense (Korean Isolate, HY970328M)

  • Ki, Jang-Seu;Han, Myung-Soo
    • Ocean Science Journal
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    • 제40권3호
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    • pp.155-166
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    • 2005
  • New PCR primers (N=18) were designed for the isolation of complete SSU to LSU rDNA sequences from the dinoflagellate Alexandrium tamarense. Standard PCR, employing each primer set selected for amplifications of less than 1.5 kb, successfully amplified the expected rDNA regions of A. tamarense (Korean isolate, HY970328M). Complete SSU, LSU rDNAs and ITS sequences, including 5.8S rDNA, were recorded at 1,800 bp, 520 bp and 3,393 bp, respectively. The LSU rDNA sequence was the first report in Alexandrium genus. No intron was found in the LSU rRNA coding region. Twelve D-domains within the LSU rDNA were put together into 1,879 bp (44.4% G+C), and cores into 1514 bp (42.8% G+C). The core sequence was significantly different (0.0867 of genetic distance, 91% sequence similarity) in comparison with Prorocentrum micans (GenBank access. no. X16108). The D2 region was the longest in length (300 bp) and highly variable among the 12 D-domains. In a phylogenetic analysis using complete LSU rDNA sequences of a variety of phytoplankton, A. tamarense was clearly separated with high resolution against other species. The result suggests that the sequence may resolve the taxonomic ambiguities of Alexandrium genus, particularly of the tamarensis complex.

Keratitis by Acanthamoeba triangularis: Report of Cases and Characterization of Isolates

  • Xuan, Ying-Hua;Chung, Byung-Suk;Hong, Yeon-Chul;Kong, Hyun-Hee;Hahn, Tae-Won;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • 제46권3호
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    • pp.157-164
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    • 2008
  • Three Acanthamoeba isolates (KA/E9, KA/E17, and KA/E23) from patients with keratitis were identified as Acanthamoeba triangularis by analysis of their molecular characteristics, a species not previously recognized to be a corneal pathogen. Epidemiologic significance of A. triangularis as a keratopathogen in Korea has been discussed. Morphologic features of Acanthamoeba cysts were examined under a microscope with differential interference contrast (DIC) optics. Mitochondrial DNA (mtDNA) of the ocular isolates KA/E9, KA/E17, and KA/E23 were digested with restriction enzymes, and the restriction patterns were compared with those of reference strains. Complete nuclear 188 and mitochondrial (mt) 16S rDNA sequences were subjected to phylogenetic analysis and species identification. mtDNA RFLP of 3 isolates showed very similar patterns to those of SH621, the type strain of A. triangularis. 16S and 18S rDNA sequence analysis confirmed 3 isolates to be A. triangularis. 18S rDNA sequence differences of the isolates were 1.3% to 1.6% and those of 16S rDNA, 0.4% to 0.9% from A. triangularis SH621. To the best of our knowledge, this is the first report, confirmed by 18S and 16S rDNA sequence analysis, of keratitis caused by A. triangularis of which the type strain was isolated from human feces. Six isolates of A. triangularis had been reported from contaminated contact lens cases in southeastern Korea.

Variation in the number of nucleoli and incomplete homogenization of 18S ribosomal DNA sequences in leaf cells of the cultivated Oriental ginseng (Panax ginseng Meyer)

  • Chelomina, Galina N.;Rozhkovan, Konstantin V.;Voronova, Anastasia N.;Burundukova, Olga L.;Muzarok, Tamara I.;Zhuravlev, Yuri N.
    • Journal of Ginseng Research
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    • 제40권2호
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    • pp.176-184
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    • 2016
  • Background: Wild ginseng, Panax ginseng Meyer, is an endangered species of medicinal plants. In the present study, we analyzed variations within the ribosomal DNA (rDNA) cluster to gain insight into the genetic diversity of the Oriental ginseng, P. ginseng, at artificial plant cultivation. Methods: The roots of wild P. ginseng plants were sampled from a nonprotected natural population of the Russian Far East. The slides were prepared from leaf tissues using the squash technique for cytogenetic analysis. The 18S rDNA sequences were cloned and sequenced. The distribution of nucleotide diversity, recombination events, and interspecific phylogenies for the total 18S rDNA sequence data set was also examined. Results: In mesophyll cells, mononucleolar nuclei were estimated to be dominant (75.7%), while the remaining nuclei contained two to four nucleoli. Among the analyzed 18S rDNA clones, 20% were identical to the 18S rDNA sequence of P. ginseng from Japan, and other clones differed in one to six substitutions. The nucleotide polymorphism was more expressed at the positions 440-640 bp, and distributed in variable regions, expansion segments, and conservative elements of core structure. The phylogenetic analysis confirmed conspecificity of ginseng plants cultivated in different regions, with two fixed mutations between P. ginseng and other species. Conclusion: This study identified the evidences of the intragenomic nucleotide polymorphism in the 18S rDNA sequences of P. ginseng. These data suggest that, in cultivated plants, the observed genome instability may influence the synthesis of biologically active compounds, which are widely used in traditional medicine.