• Title/Summary/Keyword: ribosomal DNA.

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Molecular Analysis of Exophiala Species Using Molecular Markers

  • Chee, Hee-Youn;Kim, Yoon-Kyoung
    • Mycobiology
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    • v.30 no.1
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    • pp.1-4
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    • 2002
  • Genetic relatedness of medically important Exophiala species such as E. dermatitidis, E. mansonii, and three E. jeanselmei varieties: jeanselmei, lecanii-corni, and heteromorpha was examined using PCR-RFLP(restriction fragment length polymorphism) of ribosomal DNA, M-13, $(GTG)_5$ and nucleotide sequences of ribosomal ITS(internal transcribed space) II regions. Three E. jeanselmei varieties showing distinct band patterns for each DNA markers as well as different nucleotide sequences of ribosomal ITS II regions could be considered as a separate species. E. dermatitidis and E. mansonii demonstrated the identical band patterns of RFLP of ribosomal DNA, M-13, and $(GTG)_5$ markers. However, nucleotides sequences of ribosomal ITS II region were different between these two species.

The Base Sequence of ITS and Genetic Variation in Sarcodon Aspratus (능이버섯의 ITS염기서열과 유전적 변이)

  • Kim Jong Bong
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.963-966
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    • 2004
  • The sequence of ITS (partial 16S ribosomal DNA, complete ITS1, 5.8S ribosomal DNA and ITS2, and partial 28S ribosomal DNA) was analysed by PCR and autosequencing in Sarcodon aspratus. The ITS lenght of S. aspratus was 716 base pair. As this sequence compared with other reports on S. aspratus (ace No AF335110), the sequence variation based on nucleotide deletion and substitution was $1.8\%$. This nucleotide variation rate in same species was very higher than in other species. Also, the sequence varitation rates between this S. aspratus and S. imbricatus, and S. squamus were $8\%\;and\;10\%$, respectively. This results suggested that the high sequence variation of S. aspratus might be caused specific host and inhabitat environment which limited gene flow.

Differentiation of Intraspecific Groups within isolates of Rhizoctonia solani Using PCR-RFLP of Ribosomal DNA (Ribosomal DNA의 PCR-RFLP에 의한 국내산 Rhizoctonia solani 균주들의 종내그룹의 구분)

  • 홍승범;고승주;류진창;김완규;김인수
    • Korean Journal Plant Pathology
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    • v.14 no.2
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    • pp.157-163
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    • 1998
  • Genetic diversity among 27 isolates of Rhizoctonia solani, which were obtained from diseased crops in Korea and classified into 9 intraspecific groups by anastomosis test and cultural characteristics, was studied by PCR-RFLP. Gene regions of nuclear 17S ribosomal DNA and internal transcribed spacers including 5.8S rDNA of the isolates were amplified with polymerase chain reaction and digested with 12 restriction enzymes. Differences of restriction patterns were not shown among isolates within each intraspecific groups, however, each anastomosis group and culturala type sowed unique restriction fragment length polymorphisms by restriction patterns using HaeIII, Cfr13I and MspI. The results suggest that PCR-FRLP of rDNA using three restriction enzymes could be used to differentiate intraspecific groups of Rhizoctonia solani in Korea.

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Cloning and Sequence Analysis of Ribosomal Protein S4 cDNA from Root of Panax ginseng

  • In Jun-Gyo;Lee Bum-Soo;Song Won-Seob;Bae Chang-Hyu;Choi Seong-Kyu;Yang Deok-Chun
    • Plant Resources
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    • v.8 no.2
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    • pp.110-115
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    • 2005
  • Ribosomal protein complex with ribosomal RNA to form the subunits of the ribosome serve essential functions in protein synthesis. A full-length cDNA (PRPS4) encoding ribosomal protein S4 has been isolated and its nucleotide sequence determined in ginseng plant (Panax ginseng). A PRPS4 cDNA is 1105 nucleotides long and has an open reading frame of 792 bp with a deduced amino acid sequence of 264 residues (pI 10.67). The deduced amino acid sequence of PRPS4 matched to the previously reported ribosomal protein S4 genes. Their degree of amino acid identity ranged from 68 to $92\%$. Phylogenetic analysis based on the amino acid residues showed that the PRPS4 grouped with ribosomal protein S4 of S. tuberosum (CAA54095).

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Cloning of 17S-Ribosomal RNA Gene from the Hygromycin Resistant Tetrahymena thermophila (Hygromycin내성 Tetrahymena thermophila의 17S-Ribosomal RNA유전자의 Cloning)

  • 홍용기
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.133-137
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    • 1986
  • 17S-ribosomal RNA gene from the hygromycin resistant protozoan Tetrahymena thermophila hmr 3 was cloned on E. coli vector pBR 322 as part of study to work the 17S-rRNA structure and the mechanism of hygromycin resistance. The 17S-rDNA was inserted into the Hind 111 site of pBR 322. The clones having recombinant plasmid were selected by the method of colony hybridization with a 17S-rDNA probe of wild type B1868. The orientation of 17S-rDNA insert was located near the tetracycline resistant gene of pBR 322 in a clone 5-19 with the recombinant plasmid.

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Cloning and Regulation of Schizosaccharomyces pombe Gene Encoding Ribosomal Protein S20

  • Lee, Yoon-Jong;Kim, Kyunghoon;Park, Eun-Hee;Ahn, Ki-Sup;Kim, Daemyung;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.31-36
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    • 2001
  • A cDNA clone encoding the ribosomal protein S20 has been isolated from the Schizosaccharomyces pombe cDNA library by colony hybridization. The insert contained in the original plasmid pYJ10 was transferred intro shuttle vector pRS316 generate plasmid pYJll. The dDNA insert of plasmid pYJll, contains 484 nucleotides and encodes a protein of 118 amino acids with a calculated mass of 13,544 daltons. The deduced amino acid sequence of S. pombe ribosomal protein S20 is very homologous with fruit fly, rat, and budding yeast counterparts. It is also homologous with Xenopus S22 ribosomal protein. S. pombe ribosomal protein S20 appears to be relatively hydruphobic except the C-terminal region. The 728 bp upstream region of the S20 gene was amplified from chromosomal DNA and transferred into the BamHI/EcoRI site of the promoterles $\beta$-galactosidase gene of the vector YEp357R, which resulted in fusion plasmid pYS20. The synthesis of $\beta$-galactosidase from the fusion plasmid appeared to be the highest in the mid-exponential phase. The S. pombe cells with the fusion plasmid grown at 35$\^{C}$ gave lower $\beta$-galactosidase activity than the cells grown at 30$\^{C}$. Computer analysis showed the consensus sequence CAGTCACA in the upstream regions of various ribosomal protein genes in S. pombe, which would be involved in the coordinated expression of small ribosomal proteins.

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Phylogenetic relationship of ribosomal ITS2 and mitochondrial COI among diploid and triploid Paragonimus westermani isolates

  • Park, Gab-Man;Im, Kyung-Il;Yong, Tai-Soon
    • Parasites, Hosts and Diseases
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    • v.41 no.1
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    • pp.47-55
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    • 2003
  • We compared patterns of intraspecific polymorphism of two markers with contrasting modes of evolution, nuclear ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA), in the lung fluke, diploid and triploid Paragonimus westermani from three geographical regions of Korea. The genetic distances between three populations of Korean diploid and triploid P. westermani showed no significant difference in the nucleotide sequences of the mitochondrial cytochrome c oxidase subunit 1 (mtCOI) and ribosomaal second internal transcribed spacer (ITS2) genes. A highly resolved strict-consensus tree was obtained that illustrated phylogenetically useful information of the ITS2 and mtCOI sequences from diploid and triploid P. westermani.

General properties and phylogenetic utilities of nuclear ribosomal DNA and mitochondrial DNA commonly used in molecular systematics

  • Hwang, Ui-Wook;Kim, Won
    • Parasites, Hosts and Diseases
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    • v.37 no.4
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    • pp.215-228
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    • 1999
  • To choose one or more appropriate molecular markers or gene regions for resolving a particular systematic question among the organisms at a certain categorical level is still a very difficult process. The primary goal of this review, therefore, is to provide a theoretical information in choosing one or more molecular markers or gene regions by illustrating general properties and phylogenetic utilities of nuclear ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) that have been most commonly used for phylogenetic researches. The highly conserved molecular markers and/or gene regions are useful for investigating phylogenetic relationships at higher categorical levels (deep branches of evolutionary history). On the other hand, the hypervariable molecular markers and/or gene regions are useful for elucidating phylogenetic relationships at lower categorical levels (recently diverged branches). In summary, different selective forces have led to the evolution of various molecular markers or gene regions with varying degrees of sequence conservation. Thus, appropriate molecular markers or gene regions should be chosen with even greater caution to deduce true phylogenetic relationships over a broad taxonomic spectrum.

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Isolation and Characterization of the Ribosomal Protein 46 Gene in Drosophila melanogaster

    • Animal cells and systems
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    • v.2 no.1
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    • pp.113-116
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    • 1998
  • A cDNA clone coding for ribosomal protein 46 (rp46) which is a component of 60S ribosomal large subunit has been identified from Drosophila melanogaster. A cDNA clone encoding S. cerevisiae rp46 was used as a probe to screen a Drosophila larvae cDNA library. The DNA sequence analysis revealed that the cDNA coding for Drosophils rp46 contains a complete reading frame of 153 nucleotides coding for 51 amino acids. The deduced amino acid sequence showed 71-75% homology with those of other eukaryotic organisms. Northern blot analysis showed that about 1-kb rp46 transcripts are abundant throughout fly development. Whole mount embryonic mRNA in situ hybridization also showed no preferential distribution of the transcripts to any specific region. The chromosomal in situ hybridization revealed that the identified gene is localized at position 60C on the right arm of the second polytene chromosome with a possibility of single copy.

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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.