• 제목/요약/키워드: 18S ribosomal RNA

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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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    • 제21권3호
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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.

토마토생장촉진효과가있는 Paraburkholderia phenoliruptrix T36S-14 균주의유전체염기서열 (Complete Genome Sequence of Paraburkholderia phenoliruptrix T36S-14, a Plant Growth Promoting Bacterium on Tomato (Solanum lycopersicum L.) Seedlings)

  • 김지원;진용주;이민주;박동석;송재경
    • 한국미생물·생명공학회지
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    • 제52권2호
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    • pp.195-199
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    • 2024
  • Paraburkholderia phenoliruptrix T36S-14, identified as a potential plant growth-promoting bacterium, was isolated from the core microbiome of tomato rhizosphere soil. When assessed for its growth promotion, Strain T36S-14 demonstrated a notable 20% increase in the fresh weight of tomato seedlings. The strain possesses two circular chromosomes, one of 4,104,520 base pair (bp) (CP119873) and the other of 3,258,072 bp (CP119874), both exhibiting G+C contents of 63.5% and 62.7%, respectively. The chromosome comprises 6,319 protein-coding sequences, 65 transfer RNA genes, and 18 ribosomal RNA genes (5S: 6, 16S: 6, and 23S: 6). Additionally, P. phenoliruptrix T36S-14 produces siderophores that promote plant growth.

TATA box binding protein and ribosomal protein 4 are suitable reference genes for normalization during quantitative polymerase chain reaction study in bovine mesenchymal stem cells

  • Jang, Si-Jung;Jeon, Ryoung-Hoon;Kim, Hwan-Deuk;Hwang, Jong-Chan;Lee, Hyeon-Jeong;Bae, Seul-Gi;Lee, Sung-Lim;Rho, Gyu-Jin;Kim, Seung-Joon;Lee, Won-Jae
    • Asian-Australasian Journal of Animal Sciences
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    • 제33권12호
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    • pp.2021-2030
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    • 2020
  • Objective: Quantitative polymerase chain reaction (qPCR) has been extensively used in the field of mesenchymal stem cell (MSC) research to elucidate their characteristics and clinical potential by normalization of target genes against reference genes (RGs), which are believed to be stably expressed irrespective of various experimental conditions. However, the expression of RGs is also variable depending on the experimental conditions, which may lead to false or contradictory conclusions upon normalization. Due to the current lack of information for a clear list of stable RGs in bovine MSCs, we conducted this study to identify suitable RGs in bovine MSCs. Methods: The cycle threshold values of ten traditionally used RGs (18S ribosomal RNA [18S], beta-2-microglobulin [B2M], H2A histone family, member Z [H2A], peptidylprolyl isomerase A [PPIA], ribosomal protein 4 [RPL4], succinate dehydrogenase complex, subunit A [SDHA], beta actin [ACTB], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], TATA box binding protein [TBP], and hypoxanthine phosphoribosyltrasnfrase1 [HPRT1]) in bovine bone marrow-derived MSCs (bBMMSCs) were validated for their stabilities using three types of RG evaluation algorithms (geNorm, Normfinder, and Bestkeeper). The effect of validated RGs was then verified by normalization of lineage-specific genes (fatty acid binding protein 4 [FABP4] and osteonectin [ON]) expressions during differentiations of bBMMSCs or POU class 5 homeobox 1 (OCT4) expression between bBMMSCs and dermal skins. Results: Based on the results obtained for the three most stable RGs from geNorm (TBP, RPL4, and H2A), Normfinder (TBP, RPL4, and SDHA), and Bestkeeper (TBP, RPL4, and SDHA), it was comprehensively determined that TBP and RPL4 were the most stable RGs in bBMMSCs. However, traditional RGs were suggested to be the least stable (18S) or moderately stable (GAPDH and ACTB) in bBMMSCs. Normalization of FABP4 or ON against TBP, RPL4, and 18S presented significant differences during differentiation of bBMMSCs. However, although significantly low expression of OCT4 was detected in dermal skins compared to that in bBMMSCs when TBP and RPL4 were used in normalization, normalization against 18S exhibited no significance. Conclusion: This study proposes that TBP and RPL4 were suitable as stable RGs for qPCR study in bovine MSCs.

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.

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.

충청지역의 임상검체로부터 분리된 대장균에 Aminoglycoside-Modifying Enzymes 확산 (Spreading of Aminoglycoside-Modifying Enzymes among Escherichia coli Isolated from Clinical Specimens in Chungcheong Province)

  • 성지연;권필승
    • 대한임상검사과학회지
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    • 제52권2호
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    • pp.136-142
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    • 2020
  • 세균의 항균제 내성율은 지난 몇십년 동안 지속적으로 상승하였으며 mobile genetic elements를 통한 항균제내성인자들의 전파는 다제내성세균의 출현 및 확산을 가중시켰다. 본연구에서는 임상검체에서 분리된 aminoglycoside에 비감수성 대장균 33주를 대상으로 mobile genetic elements를 통해 전파될 수 있는 aminoglycoside 내성인자를 조사하였다. 16S ribosomal RNA methyltransferases (RMTases)와 aminoglycoside-modifying enzyme (AME)유전자가 PCR과 DNA 염기서열분석을 통해 검출되었다. 그 결과 aac(3')-II 유전자(54.5%)를 포함하고 있는 균주가 제일 많았으며 그 다음으로 aph(3')-Ia 유전자(18.2%)가 많았고 aac(6')-Ib 유전자(15.2%)를 포함하는 균주도 있었다. RMTase 유전자는 본 연구에서는 검출되지 않았다. aac(3')-II 유전자를 포함하고 있는 18균주 중 17균주가 gentamicin에 내성을 보였으며 이중 16균주는 tobramycin에도 내성을 보였다. aac(6')-Ib 유전자를 포함하고 있는 5균주는 모두 tobramycin에 내성을 보였다. 본 연구에서 AME 유전자를 획득하는 것은 사람에서 분리된 대장균이 aminoglycoside에 내성을 나타내는 중요한 기전 중 하나임이 확인되었다. 사람으로부터 분리된 세균을 대상으로 지속적으로 항균제 내성인자를 조사하는 것은 내성세균의 확산을 막는데 필요할 것으로 사료된다.

Prevalence and Identification of Cryptosporidium spp. from Swine Slurry

  • Chun, Kae-Shik
    • 한국환경보건학회지
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    • 제35권3호
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    • pp.187-190
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    • 2009
  • Cryptosporidium spp. were detected in 17 of 135 swine lagoon samples from five farms by 18S ribosomal DNA locus and PCR. Seventeen positive samples identified were included two distinctive genotypes C. suis and Cryptosporidium sp. based on a small-subunit rRNA gene-based PCR-restriction fragment length polymorphism analysis. Cryptosporidium spp. were detected out of farrowing, farrowing and nursery (mix), and finishing. Prevalence rate was 12.6% with infection rates between 3.7 and 18.5%. We concluded that Cryptosporidium oocysts can persist in treated lagoon and potentially contaminate surface water through improper discharge. This study was undertaken for the evaluation of the infection status of the genotypes of Cryptosporidium spp. in swine lagoon.

Taxonomic Revision of Notohymena gangwonensis (Protozoa: Ciliophora), with Notes on Its Cortical Granules and Scanning Electron Micrographs

  • Moon, Ji Hye;Kim, Kang-San;Chae, Kyu Seok;Min, Gi-Sik;Jung, Jae-Ho
    • Animal Systematics, Evolution and Diversity
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    • 제36권2호
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    • pp.113-122
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    • 2020
  • From a moss sample, we isolated and identified Notohymena gangwonensis Kim et al., 2019 based on morphological and molecular data. The moss and type population has completely identical 18S rRNA (nuclear small subunit ribosomal RNA) gene sequences and both are highly similar in morphological and morphometric attributes, except for the diameter and arrangement of the cortical granules. Thus, we reexamined the type materials(i.e., micrographs and gDNA) and resulted in finding mistakes made by the authors of the species. Based on these data and supporting materials newly obtained (i.e., internal transcribed spacer [ITS] 1, ITS2, 5.8S, and partial 28S rDNA sequences, and scanning electron micrographs), we provide improved diagnosis of the species to clarify its identity. In addition, a key for Notohymena species is provided.

The description of Haematococcus privus sp. nov. (Chlorophyceae, Chlamydomonadales) from North America

  • Mark A. Buchheim;Ashley Silver;Haley Johnson;Richard Portman;Matthew B. Toomey
    • ALGAE
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    • 제38권1호
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    • pp.1-22
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    • 2023
  • An enormous body of research is focused on finding ways to commercialize carotenoids produced by the unicellular green alga, Haematococcus, often without the benefit of a sound phylogenetic assessment. Evidence of cryptic diversity in the genus means that comparing results of pigment studies may be confounded by the absence of a phylogenetic framework. Moreover, previous work has identified unnamed strains that are likely candidates for species status. We reconstructed the phylogeny of an expanded sampling of Haematococcus isolates utilizing data from nuclear ribosomal markers (18S rRNA gene, 26S rRNA gene, internal transcribed spacer [ITS]-1, 5.8S rRNA gene, and ITS-2) and the rbcL gene. In addition, we gathered morphological, ultrastructural and pigment data from key isolates of Haematococcus. Our expanded data and taxon sampling support the concept of a new species, H. privus, found exclusively in North America. Despite overlap in numerous morphological traits, results indicate that ratios of protoplast length to width and akinete diameter may be useful for discriminating Haematococcus lineages. High growth rate and robust astaxanthin yield indicate that H. rubicundus (SAG 34-1c) is worthy of additional scrutiny as a pigment source. With the description of H. privus, the evidence supports the existence of at least five, species-level lineages in the genus. Our phylogenetic assessment provides the tools to frame future pigment investigations of Haematococcus in an updated evolutionary context. In addition, our investigation highlighted open questions regarding polyploidy and sexuality in Haematococcus which demonstrate that much remains to be discovered about this green flagellate.

인삼 뿌리썩음병균에 항균활성이 있는 방선균 BK185의 분리 및 특성 (Isolation and Characterization of Actinomycete Strain BK185 Possessing Antifungal Activity against Ginseng Root Rot Pathogens)

  • 김병용;배문형;안재형;원항연;김성일;김완규;오동찬;송재경
    • 농약과학회지
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    • 제18권4호
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    • pp.396-403
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    • 2014
  • 인삼은 한국을 포함한 여러 국가에서 경제적으로 중요한 약용작물이다. 인삼에서 흔히 발생하는 뿌리썩음 병균을 친환경적으로 방제하기 위해서 병원균에 대해서 항균성이 있는 미생물을 탐색하였다. 토양에서 분리한 방선균 BK185균주는 인삼에 뿌리썩음병을 일으키는 병원균들(Cylindrocarpon, Fusarium, Rhizoctonia, Sclerotinia)에 대해서 높은 항균 활성을 보였다. 선발 균주 BK185의 동정을 위해서 16S rRNA 유전자 염기서열을 분석한 결과 Streptomyces 속에 해당하는 것으로 분석되었다. 특히, S. sporoclivatus 및 S. geldanamycininus와 99.6% 이상으로 매우 높은 유사도를 보였다. 선발 균주가 생산하는 2차 대사물질을 예측하기 위해서, 생산에 관여하는 생합성 유전자인PKS (Type-I polyketide synthase)와 NRPS (Non-ribosomal polypeptide synthetase) 유전자를 PCR반응을 통해 검출하였다. 검출된 유전자는 클로닝을 통해 염기서열을 결정하였다. 또한 최적 배양조건하에서 생산된 대사물질을 LC/MS로 분석하였고, geldanamycin 계열의 항생물질이 생산됨을 확인하였다.