• Title/Summary/Keyword: ssu rDNA

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Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum

  • Yu, Jae-Ran;Lee, Soo-Ung;Park, Woo-Yoon
    • Parasites, Hosts and Diseases
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    • 제47권3호
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    • pp.293-297
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    • 2009
  • Improved methods for detection of Cryptosporidium oocysts in environmental and clinical samples are urgently needed to improve detection of cryptosporidiosis. We compared the sensitivity of 7 PCR primer sets for detection of Cryptosporidium parvum. Each target gene was amplified by PCR or nested PCR with serially diluted DNA extracted from purified C. parvum oocysts. The target genes included Cryptosporidium oocyst wall protein (COWP), small subunit ribosomal RNA (SSU rRNA), and random amplified polymorphic DNA. The detection limit of the PCR method ranged from $10^3$ to $10^4$ oocysts, and the nested PCR method was able to detect $10^0$ to $10^2$ oocysts. A second-round amplification of target genes showed that the nested primer set specific for the COWP gene proved to be the most sensitive one compared to the other primer sets tested in this study and would therefore be useful for the detection of C. parvum.

Molecular Detection of Harmful Dinoflagellates (Dinophyceae) in Ballast Water (선박평형 수 내 유해 와편모조류(Dinophyceae)의 분자생물학적 검출)

  • Park, Tae-Gyu;Kim, Sung-Yeon
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • 제15권1호
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    • pp.36-40
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    • 2010
  • Ballast water has been known as a major vector for global dispersal of toxic dinoflagellates and other microalgae. In this study, biodiversity in ships’ ballast water was examined using a dinoflagellate-oriented PCR primer set and species-specific real-time PCR. While motile dinoflagellates could be observe at very low cell densities by light microscopy,a wide range of dinoflagellate taxa including parasitic and phototrophic pico-dinoflagellates as well as harmful species to marine fish/shellfish was detected when techniques for cloning/sequencing of SSU rDNA of sample cells were used. Present result suggests that molecular methods including species-specific PCR primers may offer rapid and accurate detection of invasive species in ballast water.

PCR and RFLP variation of conserved region of small subunit ribosomal DNA among Acanthamoeba isolates assigned to either A. castellanii or A. polyphaga (카스텔라니가시아메바 혹은 대식가시아메바로 분류된 분리주간의 ribosomaIDNA conserved region의 PCR-RFLP의 다양성)

  • 공현희;정동일
    • Parasites, Hosts and Diseases
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    • 제34권2호
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    • pp.127-134
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    • 1996
  • Twelve isolates of Accnthamoebc app. assigned to either A. castellanii or A. poIMphoSa, and type strains of A. culbefsoni, A. henIWi, A. pqkestinefiE, and A. astronyxi,s were examined by restriction fragment length polymorphism (RFLP) of a conserved region of small subunit ribosomal RMh gene (ssu rDNA) amplified by polymerase chain reaction (PCR). The PCR products of the isolates measured approximately 910-930 bp, except for that of A. astronyxis which was extraordinarily long, approximately 1,170 bp. Average of estimated sequence divergence of the amplified DNA among the isolates assigned to A. castellanii was 9.8% whereas that among the isolates assigned to A. polvphusn 9.6%. The maximum intraspecific sequence divergence among the isolates assigned to A. costellanii was observed between the Chang and Ma strains (17.3%) while that among the isolates assigned to A. poIWphosa was observed between KA/S3 and KA/S7 strains (16.1%). The both maximum sequence divergences were much greater than the minimum interspecific sequence divergence between A. cnstellnnii and A. polwphasa (2.6%) which appeared between the Castellani (or CCAP 1501/12 g) and KA/S3 strains. The PCR-RFLP patterns of A. culbertsoni, A. healyi, A. palestinensis, and A. ostronvxis were quite diverse from one another and from those of isolates assigned to either A. castellanii or A. polyphoga. It is suggested that taxonomic validity of the isolates assigned to either A. castellnnii or A. polyphoga should be reevaluated.

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Phylogenetics of Trichaptum Based on Mitochondrial Small Subunit rDNA Sequences

  • Ko, Kwan-Soo;Jung, Hack-Sung
    • Journal of Microbiology
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    • 제35권4호
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    • pp.259-263
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    • 1997
  • To study the phylogenetic relationships of species of Trichaptum and to infer intraspecific dibergence of T. abietinum, partial mitochondrial small subunit rDNA sequences were determined. Six strains of T. abietinum, two of T. biforme, and one of T. fusco-violaceum were examined. Parsimony and distance analyses showed that each Trichaptum species forms a distinct group and that T. abietinum consists of two or more subgroups. Strains from North America were distantly related to one another but the European strain formed an independent group with three Korean strains, suggesting the possibility that Korean taxa may be phylogenetically closer to European taxa than to North American taxa.

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Nuclear SSU and Plastid rbcL Genes and Ultrastructure of Mallomonas caudata (Synurophyceae) from Korea (한국산 Mallomonas caudata (Synurophyceae)의 미세구조, 핵 SSU 그리고 색소체 rbcL 유전자)

  • Kim, Han-Soon;Shin, Woong-Ghi;Boo, Sung-Min
    • Korean Journal of Ecology and Environment
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    • 제40권3호
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    • pp.387-394
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    • 2007
  • Despite geographic barriers such as oceans, many freshwater algal species inhabit different continents of the world. A unicellular freshwater alga, Mallomonas caudeata, commonly occurring in Asia, Europe, and America of the northern Hemisphere, is closely related to human life such as monitoring blooms and defecting changes in climates. In order to demonstrate its occurrence in Korea and to infer its phylogeny, we sequenced nuclear SSU and plastid rbcL genes from isolates collected in six different reservoirs. We have also investigated transmission electron microscopy of the Korean isolates. SSU sequences of the species from Korea and USA were almost identical, having pair-wise divergences of 0.06% in SSU and 0.45% in rbcL. Both gene trees revealed that the species was clearly separated from other species of the genus, while the genus was not monophyletic. Rhizoplasts are composed of microfibrils organised in striated rootlets attached to the multilayered plate of basal bodies and arranged on the surface of the nucleus at their distal ends. The rhizoplast constitutes a basal body-nucleus connector similar to that of typical Synurophyceas. The results that Mallomonas was not supported by both SSU and rbeL data sets require a further study with additional taxon sampling.

Acaulospora jejuensis, a New Species in Glomeromycota from Korea

  • Park, Hyeok;Ka, Kang-Hyeon;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • 제49권4호
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    • pp.425-431
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    • 2021
  • New species of arbuscular mycorrhizal fungi (Glomeromycota), Acaulospora jejuensis, was isolated from rhizosphere soils of Miscanthus sinensis in the grassland in Jeju Island of Korea. The species was identified using the morphological characteristics of the spores and the molecular phylogenetic analysis using partial DNA sequences from small subunit rDNA (SSU), internal transcribed spacer (ITS), and large subunit rDNA (LSU). Phylogenetic analysis placed the fungal species in a distinct clade within genus Acaulospora. Also, the species exhibited the morphological characteristics distinct from the other members of the genus. Therefore, Acaulospora jejuensis was described as a novel species from Korea.

Genetic Diversity and Pathogenicity of Cylindrocarpon destructans Isolates Obtained from Korean Panax ginseng

  • Song, Jeong Young;Seo, Mun Won;Kim, Sun Ick;Nam, Myeong Hyeon;Lim, Hyoun Sub;Kim, Hong Gi
    • Mycobiology
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    • 제42권2호
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    • pp.174-180
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    • 2014
  • We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.

Restriction Fragment Length Ploymorphism of PCR Amplified Ribosomal DNA Among Korean Isolates of Phytophthora

  • Hong, Seung-Beom;Jee, Hyeong-Jin;Lee, Seung-Im;Go, Seung-Joo
    • The Plant Pathology Journal
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    • 제15권4호
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    • pp.228-235
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    • 1999
  • Genetic diversity of ninety-five Korean isolates of Phytophthora was investigated on the basis of PCR-RFLP of ribosomal DNA. The isolates were previously identified as following fifteen species by mycological and cultural characteristics; P. boehmeriae, P. cactorum, P. cambivora, P. capsici, P. cinnamoni, P. citricola, P. citrophthora, P. cryptogea, P. drechsleri, P. erythroseptica, P. infestans, P. megasperma, P. nicotianae, P. palmivora and P. sojae. The regions of small subunit (SSU) and internal transcribed spacer (ITS) of rDNA were amplified with primer pair, NS1 and ITS4, by polymerase chain reaction (PCR) and digested with nine restriction enzymes. P. boehmeriae, P. cactorum, P. cambivora, P. capsici, P. cinnamomi, P. citricola, P. citrphthora, P. infestans, P. nicotianae and P. palmivora showed specific band patterns for each species. However, P. sojae and P. erythroseptica presented identical band patterns and P. cryptogea, P. drechsleri and P. megasperma were divided into six groups, which were not compatible with delineation of the species. A group originated from cucurbits showed distinct band patterns from other groups, but the other five groups were closely related within 96.0% similarity, forming one complex group. Consequently, Korean isolates of Phytophthora were divided into thirteen genetic groups and each group was readily differentiated by comparing digestion patterns of AvaII, HaeIII, MboI, HhaI and MspI. Therefore, PCR-RFLP of rDNA using the five enzymes can be used to differentiate or identify the Phytophthora species reported in Korea so far.

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Population analysis of the toxic dinoflagellate genus Alexandrium by novel molecular markers

  • Kim, Choong-jae;Kim, Sook-Yang;Kim, Kui-Young;Kang, Young-Sil;Kim, Hak-Gyoon;Kim, Chang-Hoon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 한국양식학회 2003년도 추계학술발표대회 논문요약집
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    • pp.134-135
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    • 2003
  • The geographic expansion of the toxic dinoflagellates genus Alexandrium has been shown to be world wide ranging. The members of the genus Alexandrium ocnstituted of 20-30 species did not show substantial differences in their morphology, which is mostly referred in the 'tamarensis species complex', except some species. Though rDNA sequences variations are very few and pseudogene types are so diverse that it is difficult to use them as the specific markers. In this study, we outlined Korean and Japanese A, tamarense and A. catenella regional isolates by phylogenetic analysis inferred from no cutting alignments of LSU rDNA D1-D2 and SSU rDNA sequences to group these regional isolates. The results were compared to RFLP patterns of PCR products targeted chloroplast DNA. Lastly screening of highly repeated microsatellite DNA which is frequently used for population analysis in eukaryotes was conducted. A. catenella regional strains identified by the sequencing of rDNA D1-D2 domain were divided into at least 3 groups of type E, CMC and Chinese type, divergence root may not be deep comparing with that of A. tamarense whose pseudogenes are very variable. Results of RFLP pattern and the phylogeny of the unknown gene targeting chloroplast showed that Korean and Japanese A. catenella regional isolates were divided into 3 types: Korean, Japanese and the third CMC types. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers was useful method for population analysis of A. catenella. Various types of satellite sequences such as 5 nucleotides repeats were obtained from A. tamarense and A. catenella. The 5 nucleotides repeats were primed at the both 3'and 5' ends, and these repeats were prominent as longer repeated motifs. This repeated DNA was intercalated as internal sequences containing various types subrepeats. It is expected that these satellite DNA would be a useful molecular population marker through detail comparison among Alexandrium regional isolates to trace their transferring pathway and to prevent their human-associated their regional extents.

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