• Title/Summary/Keyword: small subunit ribosomal RNA gene

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Cloning and Characerization of the Ribosomal RNA Gene from Gonyaulax polyedra

  • Lee, Hee-Gyun;Lee, Ji-Yeon;Lee, Dong-Hee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.515-523
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    • 2001
  • The dinoflagellates have some primitive nuclear features and are evolutionarily intermediate between prokaryotes and eukaryotes. The small subunit ribosomal RAN gene, the 5.8S ribosomal RNA gene, and the internal transcribed spacer (ITS) of Gonyaulax polyedra were cloned, and their sequences were analyzed to better understand their evolutionary position. The small subunit ribosomal RNA gene was 1,794 nt long, the large subunit ribosomal RNA gene was approximately 3,500 nt long, and the 5.8S ribosomal RNA gene was 159 nt long. The first internal transcribed spacer (ITS1) was 191 nt long, and the second internal transcribed spacer (ITS2) was 185 nt long. The intergenic spacer of the ribosomal RNA gene (IGS) was about 2,200 nt long, indicating that 5,800 nt of transcribed sequences were separated by roughly 2,200 nt of intergenic spacer. The ribosomal RNA genes were repeated many times and arranged in a head-to-tail, tandemly repeated manner. The repeating unit of ribosomal RNA gene of G. polyedra was proposed to be 8,000 nt long. Based on the lengths of ribosomal RNA, sequence alignments with representative organisms, and phylogenetic analysis on ribosomal RNA, G. polyedra appears to be one of the alveolates branched from the eukaryotic crown and, among dinoflagellates, it seems to not have emerged early.

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Identical small subunit ribosomal RNA gene nucleotide sequence of bovine Theileria isolates (Korea and Japan) and Theileria buffeli (Marula, Kenya) (한국파 일본의 소에서 분리한 Theileria 분리주와 Theiferia buffeli (Marula, Kenya)의 small subunit ribosomal RNA 유전자 염기서열의 일치)

  • 채준석;권오덕
    • Parasites, Hosts and Diseases
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    • v.36 no.1
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    • pp.47-54
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    • 1998
  • Small subunit ribosomal RNA (SSU rRNA) gene nucleotide sequences of bovine ReiLerin isolates from Korea (KLS and KCB) and japan (JHS) were determined. The genes from each isolate were amplified by the polymerase chain reaction and the approxi- mately 1.8 kb product cloned and sequenced by a modified dideoxynucleotide method. Overlapping gene segments produced with a series of primers were sequenced, resoRting in a complete DNA sequence for both forward and reverse strands of the SSU rRNA genes of each isolate. SSU rRNA gene sequences (termed Type A) were identical among the bovine ReiLeri,n isolates from Korea and the isolate from Japan. A GenBank data library homolo- gy search showed the sequence to be the same as that listed as leiLeyia buKeLi isolated from cattle in Marula, Kenya.

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Identification and sequence analysis of small subunit ribosomal RNA gene of bovine Theileria isolates from Korea and Japan (한국과 일본 소에 감염된 Theileria 분리주의 small subunit ribosomal 유전자의 동정 및 분석)

  • Chae, Joon-seok;Park, Jin-ho;Kwon, Oh-deog;Waghela, Suryakant D.;Holman, Patricia J.;Wagner, Gerald G.;Lee, Joo-mook
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.909-917
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    • 1998
  • Six different sequences types(A through E and H) and a subtype(Bl) of the small subunit ribosomal RNA(SSUrRNA) gene were found in bovine Theileria isolates from different areas of Korea and Japan. The sequences were aligned and three hypervariable regions were observed in the nucleotide position ranges 212~231, 261~270 and 632~690. Five of the Theileria isolates yielded sequence type A; these were the field isolates KCB, KCN, and KCJ, and the laboratory stock KLS, all from Korea, and a single isolate from Japan (JHS). This sequence type is identical to the SSUrRNA gene sequence listed for Theileria buffeli (GenBank Accession No. Z15106) from Marula, Kenya. The Korean field isolate KKB yielded only a single sequence type (B), but multiple sequence types were found in some isolates. For example, KCB and JHS isolates yielded both types A and B ; isolate KKW showed types B and H; isolate KCN showed types A, C, and D ; and isolate KCJ showed types A, B, E, and a subtype B1. Finding of the multiple sequences SSUrRNA gene sequences suggests that bovine Theileria isolates from both Korea and Japan may consist of mixed populations.

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The Infection of Heterosporis anguillarum in Cultured Shortfin Eel (Anguilla bicolor pacifica) (양식동남아산 뱀장어(Anguilla bicolor pacifica)의 Heterosporis anguillarum 감염)

  • Kim, Jin-Do;Do, Jeong-Wan;Choi, Hye-Sung;Jo, Hyae-In;Lee, Nam-Sil;Kim, Young-Dae
    • Korean Journal of Environmental Biology
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    • v.32 no.4
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    • pp.382-388
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    • 2014
  • Shortfin eel (Anguilla bicolor pacifica) is a species of commercial importance and its production is greatly affected due to the infection by Heterosporis anguillarum. In this study, we evaluated the effect of H. anguillarum infection on the growth of Shortfin eel. A disease that trunk muscle of cultured shortfin eel, Anguilla bicolor pacifica, were irregular and resulted in death, breakout of the commercial eel culture farm. We observed that the trunk muscle of infected eels were irregular and represented white or yellowish externally. Histopathologically, a great numbers of large or small spores and sporophorocysts were also observed in degenerated muscle layer. The cloning of specific gene of H. anguillarum, encoding small subunit ribosomal RNA (SSU-rRNA) was amplified by the polymerase chain reaction(PCR) from the muscle lesion of diseased eel. The size of clone gene is well matched with the size of small subunit ribosomal RNA of H. anguillarum and thus confirming the infection by H. anguillarum.

Analysis of Genetic Variation in the Small Subunit Ribosomal RNA Gene of Euplotes Ciliates for Developing Species Diagnostic Molecular Marker (종 식별 분자 마커 개발을 위한 섬모충류 Euplotes의 small subunit ribosomal RNA 변이성 분석)

  • Kim, Sun-Young;Kim, Se-Joo;Min, Gi-Sik;Yang, Eun-Jin;Yoo, Man-Ho;Choi, Joong-Ki
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.12 no.3
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    • pp.225-233
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    • 2007
  • To verify which loop regions of 18S rRNA gene are suitable as species-specific genetic markers in ciliates, we analyzed the genetic variation of 18S rRNA gene among 9 Euplotes species (Hypotrichia : Ciliophora). In our result, no inter-specific variation was detected from V1, V3 and V5 regions, and the length of V7 and V8 are 44 bp and 79 bp, respectively, which are too short to make genetic marker. In contrast, V2 and V4 may be good candidate segments of species-specific diagnostic molecular markers because these two regions are most variable ($1.75{\sim}20.61%$) and showed good inter-specific phylogeny. Furthermore, the sequences of V2 and V4 are 123 bp and 306 bp, respectively in length which are enough to make species-specific marker.

Cloning and Organization of the Ribosomal RNA Genes of the Mushroom Trichloma matsutake

  • Hwang, Seon-Kap;Kim, Jong-Guk
    • Journal of Microbiology and Biotechnology
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    • v.5 no.4
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    • pp.194-199
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    • 1995
  • A portion (7.4 kb) of ribosomal DNA tandem repeat unit from a genome of the mushroom T. matsutake has been cloned. A 1.75 kb EcoRI fragment was cloned first using S. cerevisiae 255 rRNA gene as a probe, and this was then used for further cloning. A chromosomal walking experiment was carried out and the upstream region of the 1.75 kb fragment was cloned using SmaI/BamHI enzyme, the size was estimated to be 5.2 kb in length. Part of the downstream region of the 1.75 kb fragment was also cloned using XbaI/BamHI enzymes. Restriction enzyme maps of three cloned DNA fragments were constructed. Northern hybridization, using total RNA of T. matsutake, and the restriction fragments of three cloned DNAs as probes, revealed that all four ribosomal RNA genes (large subunit[LSU], small subunit [SSU], 5.85 and 5S rRNA genes) are present in the cloned region. The gene organization of the rDNA are regarded as an intergenic spacer [IGS]2 (partial) - SSU rRNA - internal transcribed spacer [ITS]1 - 5.8S rRNA - ITS2 - LSU rRNA - IGS1 -5S rRNA - IG52 (partial).

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Phylogenetic Relationships of the Polyporaceae Based on Gene Sequences of Nuclear Small Subunit Ribosomal RNAs

  • Kim, Seon-Young;Jung, Hack-Sung
    • Mycobiology
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    • v.29 no.2
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    • pp.73-79
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    • 2001
  • The Polyporaceae is a chaotic mass of genera having poroid hymenophores in the Aphyllophorales. To classify the Polyporaceae into more natural groups, phylogenetic analyses were performed using nuclear small subunit ribosomal DNA sequences. Thirty-six species from the families of the Polyporaceae, the Hymenochaetaceae, the Ganodermataceae, the Corticiaceae, the Bondarzewiaceae, the Meruliaceae, the Steccherinaceae and the Lentinaceae were phylogenetically compared. By performing maximum parsimony analysis, seven phylogenetically meaningful groups were identified and discussed. The hyphal system, presence or absence of clamps, and the type of rot were found as important characters in defining the groups. Each group was phylogenetically significant enough to be a core member of each family when the Polyporaceae was split into smaller and more natural families.

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Genetic Characterization of Clinical Acanthamoeba Isolates from Japan using Nuclear and Mitochondrial Small Subunit Ribosomal RNA

  • Rahman, Md Moshiur;Yagita, Kengi;Kobayashi, Akira;Oikawa, Yosaburo;Hussein, Amjad I.A.;Matsumura, Takahiro;Tokoro, Masaharu
    • Parasites, Hosts and Diseases
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    • v.51 no.4
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    • pp.401-412
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    • 2013
  • Because of an increased number of Acanthamoeba keratitis (AK) along with associated disease burdens, medical professionals have become more aware of this pathogen in recent years. In this study, by analyzing both the nuclear 18S small subunit ribosomal RNA (18S rRNA) and mitochondrial 16S rRNA gene loci, 27 clinical Acanthamoeba strains that caused AK in Japan were classified into 3 genotypes, T3 (3 strains), T4 (23 strains), and T5 (one strain). Most haplotypes were identical to the reference haplotypes reported from all over the world, and thus no specificity of the haplotype distribution in Japan was found. The T4 sub-genotype analysis using the 16S rRNA gene locus also revealed a clear subconformation within the T4 cluster, and lead to the recognition of a new sub-genotype T4i, in addition to the previously reported sub-genotypes T4a-T4h. Furthermore, 9 out of 23 strains in the T4 genotype were identified to a specific haplotype (AF479533), which seems to be a causal haplotype of AK. While heterozygous nuclear haplotypes were observed from 2 strains, the mitochondrial haplotypes were homozygous as T4 genotype in the both strains, and suggested a possibility of nuclear hybridization (mating reproduction) between different strains in Acanthamoeba. The nuclear 18S rRNA gene and mitochondrial 16S rRNA gene loci of Acanthamoeba spp. possess different unique characteristics usable for the genotyping analyses, and those specific features could contribute to the establishment of molecular taxonomy for the species complex of Acanthamoeba.