• Title/Summary/Keyword: Small Subunit Ribosomal RNA

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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.

Investigation of Conservative Genes in 711 Prokaryotes (원핵생물 711종의 보존적 유전자 탐색)

  • Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.25 no.9
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    • pp.1007-1013
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    • 2015
  • A COG (Cluster of Orthologous Groups of proteins) algorithm was applied to detect conserved genes in 711 prokaryotes. Only COG0080 (ribosomal protein L11) was common among all the 711 prokaryotes analyzed and 58 COGs were common in more than 700 prokaryotes. Nine COGs among 58, including COG0197 (endonuclease III) and COG0088 (ribosomal protein L4), were conserved in a form of one gene per one organism. COG0008 represented 1356 genes in 709 of the prokaryotes and this was the highest number of genes among 58 COGs. Twenty-two COGs were conserved in more than 708 prokaryotes. Of these, two were transcription related, four were tRNA synthetases, eight were large ribosomal subunits, seven were small ribosomal subunits, and one was translation elongation factor. Among 58 conserved COGs in more than 700 prokaryotes, 50 (86.2%) were translation related, and four (6.9%) were transcription related, pointing to the importance of protein-synthesis in prokaryotes. Among these 58 COGs, the most conserved COG was COG0060 (isoleucyl tRNA synthetase), and the least conserved was COG0143 (methionyl tRNA synthetase). Archaea and eubacteria were discriminated in the genomic analysis by the average distance and variation in distance of common COGs. The identification of these conserved genes could be useful in basic and applied research, such as antibiotic development and cancer therapeutics.

Study on the variation of cellular physiology of Escherichia coli during high cell density cultivation using 2-dimensional gel electrophoresis

  • Yun, Sang-Seon;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.219-222
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    • 2000
  • Physiological changes of Escherichia coli during the fed-batch fermentation process were characterized in this study. Overall cellular protein samples prepared at the different stage of fermentation were separated by 2-dimensional gel electrophoresis (2-DE), and differently expressed 15 proteins, Phosphotransferase enzyme I, GroEL, Trigger factor, ${\beta}$ subunit of ATP synthase, Transcriptional regulator KDGR, Phosphoglycerate mutase 1, Inorganic pyrophosphatase, Serine Hydroxymethyl-transferase, ${\alpha}$ subunit of RNA polymerase, Elongation factor Tu, Elongation factor Ts, Tyrosine-tRNA ligase, DnaK suppressor protein, Transcriptional elongation factor, 30S ribosomal protein S6 were identified using matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). When bacterial cells grow to high cell density, and IPTG-inducible heterologous protein is produced, expression level of overall cellular proteins was decreased. According to their functions in the cell, identified proteins were classified into three groups, proteins involved in transport process, small-molecule metabolism, and synthesis and modification of macromolecules.

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Penicillium vietnamense sp. nov., the First Novel Marine Fungi Species Described from Vietnam with a Unique Conidiophore Structure and Molecular Phylogeny of Penicillium Section Charlesia

  • Nguyen, Van Duy;Pham, Thu Thuy
    • Mycobiology
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    • v.50 no.3
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    • pp.155-165
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    • 2022
  • Penicillium vietnamense sp. nov. was isolated from Nha Trang Bay, Vietnam in June 2017. It is phylogenetically distinct from the sister species of Penicillium section Charlesia series Indica based on multi-locus sequence typing results using internal transcribed spacer, large subunit ribosomal RNA, b-tubulin, calmodulin, and RNA polymerase II second largest subunit regions. It showed strong growth on Czapek yeast autolysate agar at 37 ℃, a strong acid production on Creatine sucrose agar, and produced short stipes, small vesicles, and subglobose to globose conidia delicately roughened with very short ridges. As the first novel marine fungi species described from Vietnam and discovered in a unique environment, the data could be significant for understanding the taxonomy and geographical distribution of marine fungi in tropical coastal systems such as Vietnam.

Utility of taxon-specific molecular markers for the species identification of herbarium specimens: an example from Desmarestia japonica (Phaeophyceae, Desmarestiales) in Korea

  • Lee, Sang-Rae;Lee, Eun-Young
    • Fisheries and Aquatic Sciences
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    • v.21 no.3
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    • pp.8.1-8.6
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    • 2018
  • Desmarestia japonica (Phaeophyceae, Desmarestiales) was recently established from the Japanese ligulate Desmarestia and is morphologically similar to D. ligulata. This species has been reported only from Japan. However, the taxonomic reports based on additional regional distributions are needed to clarify this taxonomic entity and its species boundaries. Because Desmarestia species have restricted distributions in Korea, we reexamined herbarium specimens of D. ligulata deposited at the National Institute of Biological Resources (South Korea). To improve the amplification efficiency of the polymerase chain reaction and avoid contamination by the DNA of other organisms, we developed taxon-specific molecular markers suitable for DNA barcoding of Desmarestia species. Nuclear ribosomal small subunit RNA (18S rDNA) and mitochondrial cytochrome c oxidase 1 (cox1) regions were selected as target DNA. As a result, both were successfully isolated from herbarium specimens of D. japonica acquired over 10 years. These molecular markers provide useful genetic information for herbarium specimens for which conventional molecular analysis is challenging.

A New Report on Three Species of Sordariomycetes Class Isolated from Soil in Korea

  • Das, Kallol;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.46 no.2
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    • pp.134-144
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    • 2018
  • Three fungal strains belonging to the class Sordariomycetes, namely BH-06, 17-039 and BE12-1, were isolated from soils in Korea and identified as Chaunopycnis alba, Myrothecium cinctum and Humicola olivacea, respectively. These species were confirmed according to their morphological characteristics and phylogenetic relationships determined based on internal transcribed spacer regions, as well as large subunit, small subunit, ${\beta}$-Tubulin and RNA polymerase II largest subunit sequences of ribosomal DNA. These three species are the first members of the Sordariomycetes reported in Korea.

New records of two pleurostomatids (Ciliophora: Litostomatea) from Korea

  • Kim, Se-Joo;Min, Gi-Sik
    • Journal of Species Research
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    • v.5 no.3
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    • pp.343-347
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    • 2016
  • Two pleurostomatid ciliates, Amphileptus litonotiformis Song, 1991 and Kentrophyllum setigerum (Quennerstedt, 1867), were collected from the coastal waters of Korea. Their morphologies are described based on live observation and protargol impregnation, and their diagnoses and morphometrics are provided. Additionally, sequence information of small subunit ribosomal RNA obtained from two species were reported.

Genetic Analysis of a Structural Motif Within the Conserved 530 Stem-Loop of Escherichia coli 16S rRNA

  • Szatkiewicz Jin P.;Cho Hyun-Dae;Ryou Sang-Mi;Kim Jong-Myung;Cunningham Philip R.;Lee Kang-Seok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.569-575
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    • 2006
  • The 530 stem-loop is a 46 nucleotide stem-loop structure found in all small-subunit ribosomal RNAs. Phylogenetic and mutational studies by others suggest the requirement for Watson-Crick interactions between the nucleotides 505-507 and 524-526 (530 pseudoknot), which are highly conserved. To examine the nature and functional significance of these interactions, a random mutagenesis experiment was conducted in which the nucleotides in the proposed pseudoknot were simultaneously mutated and functional mutants were selected and analyzed. Genetic analysis revealed that the particular nucleotide present at each position except 524 was not exclusively critical to the selection of functional mutants. It also indicated that basepairing interactions between the positions 505-507 and 524-526 were required for ribosomal function, and much weaker base-pairing interactions than those of the wild-type also allowed high ribosomal function. Our results support the hypothesis that the 530 pseudoknot structure may undergo a 'conformational switch' between folded and unfolded states during certain stages of the protein synthesis process by interacting with other ligands present in its environment.