• Title/Summary/Keyword: 18S-rDNA sequencing

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Two Human Cases Infected by the Horsehair Worm, Parachordodes sp. (Nematomorpha: Chordodidae), in Japan

  • Yamada, Minoru;Tegoshi, Tatsuya;Abe, Niichiro;Urabe, Misako
    • Parasites, Hosts and Diseases
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    • v.50 no.3
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    • pp.263-267
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    • 2012
  • The present study was performed to describe 2 human cases infected by the horsehair worm, Parachordodes sp., in Japan. Two gordiid worms were collected in the vomit and excreta of an 80-year-old woman in November 2009 in Kyoto city, and in the mouth of 1-year-old boy in December 2009 in Nara city, Japan, respectively. Both worms were males having bifurcated posterior ends and male gonads in cross sectional specimens. They were identified as Parachordodes sp. (Nematomorpha: Chordodidae) based on the characteristic morphologies of cross sections and areoles in the cuticle. DNA analysis on 18S rRNA partial sequence arrangements was also carried out and both worms were assumed to be close to the genus Paragordionus based on tree analysis, and far from Gordius sp. which has already been reported in humans in Japan. DNA sequencing of the Parachordodes worm does not appear on the database; therefore, more information on the gene sequences of the genus Parachordodes from humans, animals, or intermediates is required.

Studies on the cloning gp50 and gp63 genes of Pseudorabies virus(Shope strain) (Pseudorabies virus의 gp50과 gp63 유전자 클로닝에 관한 연구)

  • Kweon, Chang-hee;Song, Jae-young;Kim, Byoung-han;Lee, Jung-bok;Lee, jae-chin;An, Soo-hwan;Lee, Yong-soon;Susumu, Maeda
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.311-318
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    • 1991
  • The DNA fragment representing for Pseudorabies gp50 and gp60(Shope) was cloned by recombinant techniques. The viral DNA was extracted from the infected cells and digested with Bam HI. The 6.8 Kb of Bam HI fragment was isolated from agarose gel and further digested with Nde I followed by Klenow treatment. The blunt ended 4.9Kb fragment was cloned into pTZ18R plasmid vector. The upstream region of gp50 was further manipulated to remove its 5' promoter region and create EcoRl site for possible eukaryotic expression system. The result of partial sequencing of cloned DNA indicated that Shope strain showed 95% homology with gp50 of Rice strain.

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Phylogenetic Positioning of a Strongyloides stercoralis Isolate Recovered from a Korean Patient and Comparison with Other Asian Isolates

  • Bae, Jaeho;Jeong, Mi Jin;Shin, Dong hoon;Kim, Hyun Woo;Ahn, Sung Ho;Choi, Jun Ho;Yu, Hak Sun
    • Parasites, Hosts and Diseases
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    • v.58 no.6
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    • pp.689-694
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    • 2020
  • Strongyloidiasis is caused by Strongyloides stercoralis and is one of the most neglected tropical diseases in tropical and subtropical regions. Although several strongyloidiasis cases have been reported in Korea, genetic analysis of Korean isolates is still incomplete. In this study, a parasite was isolated from a 61-year-old man diagnosed with strongyloidiasis during the treatment of lymphoma on his retroperitoneal lymph node. Diffuse symmetric wall thickening from the ascending to descending colon and a nematode-infected intestine was observed following microscopic examination. Genomic DNA was isolated from a patient tissue block, and S. stercoralis was identified by PCR and sequencing (18S rDNA). In order to determine phylogenetic location of a Korean isolate (named KS1), we analyzed cox1 gene (500-bp) and compared it with that from 47 previous S. stercoralis isolates (28 human isolates and 19 canid isolates) from Asian countries. Our results showed that phylogenetic tree could clearly be divided into 5 different groups according to hosts and regions. KS1 was most closely related with the Chinese isolates in terms of genetic distance.

Molecular Identification of a Sea Anemone (Cnidaria: Anthozoa: Actiniaria) Obtained in Gijang, Busan (부산 기장에서 채집된 말미잘의 분자생물학적 방법을 이용한 동정)

  • Yoo, Sang Joon;Kim, Do-Hyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.4
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    • pp.447-452
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    • 2017
  • In this study, we tried to identify a sea anemone collected from the coast of Gijang, Busan. The anemone was morphologically similar to species belonging to the genus Anthopleura, but its morphological characteristics did not allow for confirmed identification to species level. Multiple genes from mitochondrial cytochrome oxidase III, 12S and 16S rRNA, and nuclear 18S and 28S rRNA, were amplified for multilocus sequence typing (MLST) analysis using genomic DNA extracted from the sampled anemone and a different primer set. Based on the MLST analysis, the anemone obtained in this study was identified as Anthopleura artemisia. Also, the sequence of internal transcribed spacer-2 was most closely related to A. artemisia, indicating that this single region might be useful for anemone identification. This study shows significance of molecular identification for sea anemones, and will be helpful in studies of sea anemone identification using genotyping-by-sequencing.

Isolation and Characterization of Oligotrophic Strains with High Enzyme Activity from Buckwheat Sokseongjang (메밀 속성장 유래 효소활성 우수 저영양성 균주 분리 및 특성)

  • Lee, Sung-Young;Kim, Ji-Yeun;Baek, Sung-Yeol;Yeo, Soo-Hwan;Koo, Bon-Sung;Park, Hye-Young;Choi, Hye-Sun
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.735-741
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    • 2011
  • Bealmijang is a short-term fermented regional product that is prepared with soybean and extra ingredients. In this study, starter strain candidates were screened from Bealmijang for fermented soybean paste products. Twenty one bacterial strains producing extracellular enzymes (amylase, cellulase, protease, xylanase and lipase) were isolated from Bealmijang, buckwheat sokseongjang. The isolates were assessed for fibrinolytic and antibacterial activities, and salt tolerance. Strain HJ18-4, identified as Bacillus subtilis (AB601598) by biochemical properties (89.6%) and 16S rDNA sequencing (100%), showed the highest enzymatic, fibrinolytic, and antibacterial activities among the isolates. Although the growth of HJ18-4 was inhibited by the increase of NaCl concentration, the growth still exceeded that of B. subtilis KACC 10114 at 5% and 10% NaCl. These results suggest that B. subtilis HJ18-4 is suitable as a starter for soybean paste manufacture.

Isolation and Characteristics of Photosynthetic Bacterium, Erythrobacter longus SY-46 which Produces Bacterial Carotenoids (Bacterial Carotenoids를 생산하는 광합성세균 Erythrobacter longus SY-46의 분리 및 특성)

  • Kim, Yun-Sook;Lee, Dae-Sung;Jeong, Seong-Yun;Lee, Won-Jae
    • Journal of Environmental Science International
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    • v.17 no.4
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    • pp.469-477
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    • 2008
  • The aerobic photosynthetic bacterium, which produces bacterial carotenoids was isolated and identified from coastal marine environments. This bacterium was identified by 16S rDNA sequencing and designated as Erythrobacter longus SY-46. E. longus SY-46 was Gram negative and rod shape, and the optimal culture conditions were $25^{\circ}C$, pH 7.0, and 3.0% NaCl concentration, respectively. The carbon and nitrogen sources required for the optimal growth were lactose and tryptone, respectively. Fatty acid compositions of E. longus SY-46 were $C_{18:1}$(78.32%), v-linolenic acid($C_{18:3n9.12.15c}:3.83%$), margaric acid($C_{17:0}$: 3.38%), palmitic acid($C_{16:0}$: 3.07%), and docosahexaenoic acid($C_{22:6n3}$: 2.21%). In addition, E. longus SY-46 showed the characteristic absorption peaks of bacterial carotenoids(in the region of 450 to 480 nm) and bacteriochlorophyll(770 to 772 nm). Major carotenoids of E. longus SY-46 were polyhydroxylated xanthophylls such as fucoxanthin and zeaxanthin.

Isolation and Characterization of a Marine Derived Bacterium Glaciecola sp. SL-12 Producing β-agarase (한천분해효소를 생산하는 해양유래 세균 Glaciecola sp. SL-12의 분리 및 특성)

  • Lee, Dong-Geun;Lee, Ok-Hee;Jang, Hyo-Jung;Jang, Min-Kyung;Yoo, Ki-Hwan;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.18 no.1
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    • pp.58-62
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    • 2008
  • A novel agar-degrading bacterium SL-12 was isolated from seashore of Kijang at Busan, Korea, and cultured in marine broth 2216 media. Isolated bacterium SL-12 was identified as Glaciecola genus by 16S rDNA sequencing with 98% identity. The optimum pH of the enzyme activity was 7.0 and the optimum temperature for the reaction was $30^{\circ}C$. The enzyme hydrolyzed neoagarohexaose to yield neoagarobiose as the main product, indicating that the enzyme is ${\beta}$-agarase. Thus, isolated bacterium and the enzyme would be useful for the industrial production of neoagarobiose.

Biocontrol of Late Blight and Plant Growth Promotion in Tomato Using Rhizobacterial Isolates

  • Lamsal, Kabir;Kim, Sang Woo;Kim, Yun Seok;Lee, Youn Su
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.897-904
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    • 2013
  • Seven bacterial isolates (viz., AB05, AB10, AB11, AB12, AB14, AB15, and AB17) were derived from the rhizosphere and evaluated in terms of plant growth-promoting activities and the inhibition of Phytophthora infestans affecting tomatoes in Korea. According to 16S rDNA sequencing, a majority of the isolates are members of Bacillus, and a single isolate belongs to Paenibacillus. All seven isolates inhibited P. infestans by more than 60% in vitro. However, AB15 was the most effective, inhibiting mycelial growth of the pathogen by more than 80% in vitro and suppressing disease by 74% compared with control plants under greenhouse conditions. In a PGPR assay, all of the bacterial isolates were capable of enhancing different growth parameters (shoot/root length, fresh biomass, dry matter, and chlorophyll content) in comparison with non-inoculated control plants. AB17-treated plants in particular showed the highest enhancement in fresh biomass with 18% and 26% increments in the root and shoot biomass, respectively. However, isolate AB10 showed the highest shoot and root growth with 18% and 26% increments, respectively. Moreover, the total chlorophyll content was 14%~19% higher in treated plants.

Assessment of the macroalgal diversity of Kuwait by using the Germling Emergence Method

  • Amal H. Hajiya Hasan;Dhia A. Al-Bader;Steve Woodward;Csongor Z. Antony;Jared Kok Ong;Akira F. Peters;Frithjof C. Kupper
    • ALGAE
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    • v.38 no.2
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    • pp.127-139
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    • 2023
  • Cryptic stages of diverse macroalgae present in natural substrata, "the bank of microscopic forms", were isolated into clonal cultures and identified based on both morphological characteristics and DNA barcoding. Approximately 120 clonal isolates from 308 natural substratum samples were collected from the entire coastline of Kuwait. Amongst these isolates, 77 (64%) were identified through DNA barcoding using the nuclear ribosomal small subunit, RuBisCO spacer (ITS2, tufa, rbcL, psaA, and psbA) and sequencing. Twenty-six isolates (34%) were identified in the division Chlorophyta, 18 (23%) as Phaeophyceae, and 33 (43%) as Rhodophyta. For all DNA sequences in this study, species-level cut off applied was ≥98% homology which depend entirely on the markers used. Three putative new records of Chlorophyta new for the Arabian Gulf were made: Cladophora laetevirens (Dillwyn) Kützing, Ulva torta (Mertens) Trevisan and Ulvella leptochaete (Huber) R. Nielsen, C. J. O'Kelly & B. Wysor in Nielsen, while Cladophora gracilis Kützing and Ulva ohnoi M. Hiraoka & S. Shimada are new records for Kuwait. For Phaeophyceae, Ectocarpus subulatus Kützing and Elachista stellaris Areschoug were new records for the Gulf and Kuwait. In the Rhodophyta, Acrochaetium secundatum (Lyngbye) Nägeli in Nägeli & Cramer, Ceramium affine Setchell & N. L. Gardner, Gelidium pusillum var. pakistanicum Afaq-Husain & Shameel and Dasya caraibica Børgesen are new records for the Gulf and Kuwait, while the red alga Stylonema alsidii (Zanardini) K. Drew is a new record for Kuwait. Several isolates identified corresponded to genera not previously reported in Kuwait and / or the Arabian Gulf, such as Porphyrostromium Trevisan, a new genus from the Bangiales, and two unidentified species for the Planophilaceae Škaloud & Leliaert. The isolates cultivated from substrata enhance understanding of the marine macroalgal diversity in the region and confirmed that the Germling Emergence Method is suitable for determining the actual diversity of a given study area through isolation from cryptic life-history phases.

Characterization of Newly Recorded Talaromyces veerkampii Isolated from Field Soil in Korea based on Morphology and Multigene Sequence Analysis

  • Mahesh Adhikari;Hyun Seung Kim;Hyo Bin Park;Ki Young Kim;In Kyu Lee;Eun Jeong Byeon;Ji Min Woo;Hyang Burm Lee;Youn Su Lee
    • The Korean Journal of Mycology
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    • v.50 no.4
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    • pp.347-355
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    • 2022
  • A fungal isolate belonging to the phylum Ascomycota was isolated and identified as Talaromyces veerkampii in 2017 during a survey of fungal diversity in field soils in Korea. This fungal isolate was identified and described based on macro- and micromorphological and molecular characterization. The identification was also based on partial 18S-ITS1-5.8S-ITS2-28S rDNA and calmodulin (CaM)-encoding gene sequencing data. Talaromyces veerkampii has not been previously reported in Korea. Thus, we report here a newly discovered species from soil in Korea along with its morphological and molecular characteristics.