• Title/Summary/Keyword: rDNAs

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Analysis of FISH patterns using 5S and 45S rDNAs in Codonopsis minima and C. lanceolata from Jeju Island (5S와 45S rDNA 유전자를 이용한 제주도산 애기더덕 (Codonopsis minima)과 더덕 (C. lanceolata)의 FISH 패턴 분석)

  • Kim, Soo-Young;Kim, Chan-Soo
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.3
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    • pp.186-190
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    • 2010
  • The chromosome number was identified and fluorescence in situ hybridization(FISH) mapping of 5S and 45S rDNAs were conducted for C. minima and C. lanceolata in the genus Codonopsis from Jeju island. In this study, we have confirmed that the somatic metaphase chromosome number determined as 2n=2x=16 was the same as the findings from the previous studies. While the conventional staining method makes it rather difficult to distinguish satellite chromosomes due to high degree of variability, FISH analysis produced the exact number and location of 5S and 45S rDNAs. Both species in the genus Codonopsis have a pair of 5S rDNA and their gene loci were observed on chromosome 3. Although two pairs of 45S rDNAs (one on chromosome 1 and the other on chromosome 8) were identified in both species, the 45S rDNA signals on chromosome 8 in C. minima were significantly weaker than those on chromosome 1. In addition, the 45S rDNA signals on chromosome 1 in C. lanceolata showed that the chromosome is non-homologus. In this study, we have determined cytogenetic characteristics of C. minima and C. lanceolata according to their gene replication patterns.

Physical Mapping of rDNAs Using McFISH in Anemarrhena asphodeloides Bunge (지모에서 McFISH를 이용한 rDNAs의 물리지도 작성)

  • Kim, Soo-Young;Choi, Hae-Woon;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.6
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    • pp.515-518
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    • 2004
  • Anemarrhena asphodeloides, a medicinal plant, has chromosome number of 2n=2x=22. To characterize the somatic metaphase chromosomes, physical mapping of 45S and 5S rDNAs using McFISH (multi-color fluorescence in situ hybridization) was applied. Two pairs of 45S rDNA loci were detected on the terminal regions of the short arm of chromosomes 1 and 3. A pair of 5S rDNA signal was observed on the short arm of chromosome 3. 5S rDNA site seemed to be the same locus as one of the 45S rDNA site. McFISH was very useful tool for the localization and identification of rDNAs on the metaphase chromosomes in A. asphodeloides.

Comparative Molecular Analysis of Freshwater Centric Diatoms with Particular Emphasis on the Nuclear Ribosomal DNA of Stephanodiscus (Bacillariophyceae)

  • Ki, Jang-Seu
    • ALGAE
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    • v.24 no.3
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    • pp.129-138
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    • 2009
  • DNA-based discrimination of species is a powerful way for morphologically otherwise similar species, like centric diatoms. Here, the author sequenced long-range nuclear ribosomal DNAs, spanning from the 18S to the D5 region of the 28S rDNA, of Stephanodiscus, particularly including a Korean isolate. By comparisons, high DNA similarities were detected from the rDNAs of nine Stephanodiscus (>99.4% in 18S rDNA, >98.0% in 28S rDNA). Their genetic distances, however, were significantly different (Kruskal-Wallis test, p < 0.01) compared to two related genera, namely Cyclotella and Discostella. In addition, genetic distances of 18S rDNAs were significantly different (Student’s t-test, p = 0.000) against those of the 28S rDNAs according to individual genera (Cyclotella, Discostella, and Stephanodiscus). Phylogenetic analyses showed that Stephanodiscus and Discostella showed a sister taxon relationship, and their clade was separated from a cluster of Cyclotella (1.00 PP, 100% BP). This suggests that Stephanodiscus has highly conserved sequences of both 18S and 28S rDNA; however, Stephanodiscus is well-separated from other freshwater centric diatoms, such as Cyclotella and Discostella, at the generic level.

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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    • v.18 no.5
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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.

Karyotype Analysis and rDNA Physical Mapping in Rye (Secale cereale L.) (호밀(Secale cereale L.)의 핵형분석과 rDNA의 Physical Mapping)

  • Lee, Joon Soo;Seo, Bong Bo;Kim, Min
    • Korean Journal of Breeding Science
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    • v.42 no.2
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    • pp.163-168
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    • 2010
  • This study was carried out to determine the chromosomal localization of the 5S and 18S-26S ribosomal DNA(rDNA) genes by means of fluorescence in situ hybridization(FISH) techniques, and the constitutive heterochromatin detected by means of Gimsa C-banding technique in rye(Secale cereale L.). The somatic chromosomes number was 2n=14. The karyotype consists of four pairs of metacentrics(chromosomes 1, 2, 3, and 7) and three pairs of submetacentrics(chromosomes 4, 5, and 6). Secondary constrictions appeared in the short arm of chromosome 1. The 5S rDNA genes have been located on two pairs of chromosomes 1 and 5, and 18S-26S rDNAs genes have been located on one pair of chromosome 1. 5S rDNA genes were detected on the distal region of the secondary constrictions in nucleolus organizer regions(NOR) in chromosome 1, and other detected on the intercalary region in the short arm of chromosome 5.

Molecular Systematics of the Tephritoidea (Insecta: Diptera): Phylogenetic Signal in 16S and 28S rDNAs for Inferring Relationships Among Families

  • Han, Ho-Yeon;Ro, Kyung-Eui;Choi, Deuk-Soo;Kim, Sam-Kyu
    • Animal cells and systems
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    • v.6 no.2
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    • pp.145-151
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    • 2002
  • Phylogenetic signal present in the mitochondrial 16S ribosomal RNA gene (16S rDNA) and the nuclear large subunit ribosomal RNA gene (28S rDNA) was explored to assess their utility in resolving family level relationships of the superfamily Tephritoidea. These two genes were chosen because they appear to evolve at different rates, and might contribute to resolve both shallow and deeper phylogenetic branches within a highly diversified group. For the 16S rDNA data set, the number of aligned sites was 1,258 bp, but 1,204 bp were used for analysis after excluding sites of ambiguous alignment. Among these 1,204 sites, 662 sites were variable and 450 sites were informative for parsimony analysis. For the 28S rDNA data set, the number of aligned sites was 1,102 bp, but 1,000 bp were used for analysis after excluding sites of ambiguous alignment. Among these 1000 sites, 235 sites were variable and 95 sites were informative for parsimony analysis. Our analyses suggest that: (1) while 16S rDNA is useful for resolving more recent phylogenetic divergences, 28S rDNA can be used to define much deeper phylogenetic branches; (2) the combined analysis of the 16S and 28S rDNAs enhances the overall resolution without losing phylogenetic signal from either single gene analysis; and (3) additional genes that evolve at intermediate rates between the 16S and 28S rDNAs are needed to further resolve relationships among the tephritoid families.

Karyotype Analysis and Physical Mapping of rDNAs Using McFISH in Jeffersonia dubia Benth (깽깽이풀의 핵형분석과 McFISH를 이용한 rDNA의 물리지도 작성)

  • Kim, Soo-Young;Choi, Hae-Woon;Koo, Dal-Hoe;Kim, Chan-Soo;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.1
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    • pp.48-51
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    • 2005
  • Karyotype analysis and chromosomal locailization of 45S and 5S rDNAs using McFISH (multi-color fluorescence in situ hybridization) were carried out in Jeffersonia dubia Benth., which is one of medicinal plants belonging to Berberidaceae. The somatic metaphase chromosome number was 2n=2x=12 and the size of chromosomes ranged $1.95{\sim}3.50\;{\mu}m$. The chromosome complement consisted of two pairs of metacentrics (chromosomes 1 and 3), two pairs of submetacentrics (chromosomes 2 and 4) and two pairs of subtelocentrics (chromosomes 5 and 6). In McFISH, one pair of 45S rDNA site was detected on the centromeric region of chromosome 2 and three pairs of 5S rDNA sites were detected on the short arm of chromosomes 4, 5 and 6, respectively.

Isolation and Characterization of Mitochondrial DNA from Arehicityars and Metacitrus (Archicitrus와 Metacitrus로부터 Mitochondrial DNA의 분리 및 특성)

  • 이숙영;박민희
    • The Korean Journal of Food And Nutrition
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    • v.8 no.4
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    • pp.307-317
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    • 1995
  • The purity of mtDNAs isolated from Archicitrus and Metacitrus leaves was higher in percoll density gradient centrifugation than differential and sucrose density gradient centrifugation. The most clear mtDNAs were obtained from mitochondria included in the Interface band of between 21% and 45% under isomotic, low viscosity conditions in the three step discontinuous percoll density gradient centrifugation. DNase treatment to the crude mitochondrlal suspension still more increased purity of mtDNA by the effective removal of the nuclear and chloroplast DNA and mtDNAs were appeared as a single band at middle position of tube by EtBr /cscl density gradient centrifugation. Agarose gel electrophoresis of mtDNAs resolved a single, broad band containing high molecular weight DNAs In all preparation. Yield of mtDNAs was about 110 and 2 ug Per 2009 in mature and immature leaves respectively. The mtDNA fragment patterns showed by EcoR I treatment were indistinguishable with respect to nom bet and position of bands in Archicitrus and Metacitrus. In the pattern of Hind E restriction, the Metacitrus displayed the unique band between 5.0 and 4.0kb, in addition to four fragments about 5.0, 2.4, 2.15, and 2.0kb, respectively, different from Archicitrus. Also the pattern of total mtDNAs fragment by the treatment of Pst I showed that the distinguishable fragment pat tern was not appeared in Archicitrus(C. iyo Tanaka), but about 6.0, 5.5, 5.0 and 2.Bkb fragments were appeared only in Metacitrus(C. junos Sieb). Therefore it was indicated that two species in intra-subgenus were identical each other, whereas considerable difference was revealed for inter-subgenus.

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Classification and Characterization of Bacteriophages of Lactobacillus casei -Analysis of Restriction Patterns of Phage DNA- (Lactobacillus casei bacteriophage의 분류 및 특성에 관한 연구 - Phage DNA의 제한효소 절편 비교 분석-)

  • 김영창;강현삼
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.115-121
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    • 1985
  • Five representative virulent phages (J1, TK93, K1, PD5, and CP1) and one temperate phage (.phi.1043) of Lactobacillus casei were compared to each other by analyzing the agarose gel electrophoretic patterns of restriction enzyme-digested phage DNAs. Nucleic acids of all the tested phages were double stranded DNA. DNAs of J1, TK93, K1, and ${\phi}$ 1043 phages had a size of about 42kb, but the size of PD5 and CP1 DNAs was avout 140kb. J1, TK93, K1, PD5, CP1, and ${\phi}$ 1043 DNAs were digested to 13, 13, 11, 14, 14, and 12 fragments by EcoR1, respectively, and showed its characteristec restriction patterns. Cohesive ends were present in J1, TK93, and ${\phi}$ 1043, but were absent in K1, PD5, and CP1. Restriction maps of J1 and TK93 DNAs showed nearly complete homology and their evolutionary relationship based upon the restriction analysis was discussed.

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Comparison of ITS(Internal Transcribed Spacer) and 5.8S rDNA Sequences among varieties and Cultivars in Panax ginseng

  • Yang, Deok-Chun;Yang, Key-Jin;Yoon, Eui-Soo
    • Journal of Photoscience
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    • v.8 no.2
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    • pp.55-60
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    • 2001
  • Ginseng (Panax genus) is one of the most medicinally important genera and consists of highly regarded medicines. Among the species of Panax, the ginseng species is widely known to have most medicinal quality. P. ginseng has 3 varieties, Jakyung, Chunggyung and Hwangsook, discovered in nature with different colors of stem and fruit, Jakyung has two cultivars, Yunpoong and Chunpoong. Rigorous phylogenetic analysis of these varieties and cultivars has been conducted with sequencing of rDNA region. The sequences of ITS1, ITS2 of every varieties and cultivars within P. ginseng were identical. The sequence of 5.8S rDNAs of Hwangsook variety were different from the sequences of 5.8S rDNAs of others by only one base pair at nucleotide position 14. In phylogenetic analysis and predicted RNA secondary structure study, it is assumed that evolution has proceeded from Hwangsook to other varieties. recently.

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