• Title/Summary/Keyword: 28S rDNA

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Species identification of the Anopheles kyrcanus complex found in Korea using PCR (PCR을 이용한 우리나라에서 발견되는 얼룩날개모기속 모기의 종 동정)

  • Yong, Tae-Sun;Lee, Han-Il;Lee, In-Yong;Lee, Jong-Won;Hwang, Ui-Uk
    • Journal of Korea Association of Health Promotion
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    • v.4 no.1
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    • pp.68-74
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    • 2006
  • For identification of four sibling species of the Anopheles hyrcanus complex found in Korea, the 5.8 rDNA-ITS2-28S rDNA region of each species was sequenced and the species-specific primers wee designed The amplified PCR products obtained from each species were analyzed by agarose gel electrophoresis. The result showed a single species- specific band, I.e. 559bp, 432bp, 322bp and 192bp for An. sinensis, An. sp., An. lesteri and An. pullus, respectively. In conclusion, the species-specific PCR primers designed from ITS2 variable regions functioned successfully and specifically, and can be applied as a useful tool for identifying species of the Anopheles hyrcanus complex found in Korea.

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Purification and Characterization of an Antimicrobial Substance from Bacillus subtilis HH28 Antagonistic to Bacillus cereus (Bacillus cereus를 억제하는 Bacillus subtilis HH28의 항균물질 정제와 특성규명)

  • Cha, Hyun A;Chung, Dawn;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.393-401
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    • 2014
  • A bacterium producing antimicrobial substance was isolated from cheonggukjang. The bacterium was identified as a strain of Bacillus subtilis by 16S rDNA sequencing and designated as Bacillus subtilis HH28. The antimicrobial substance produced from Bacillus subtilis HH28 was purified by 0-80% ammonium sulfate precipitation, DEAE-sepharose FF column chromatography, and Sephacryl S-200 HR gel chromatography. The molecular weight of the purified antimicrobial substance was estimated to be approximately 3,500 Da using Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis and direct detection analysis. Antimicrobial substance from B. subtilis HH28 not only inhibited B. cereus, but also Listeria monocytogenes and Vibrio parahaemolyticus. The purified antimicrobial substance was stable at $40-80^{\circ}C$, and between pH 2 and 8. Antimicrobial activity of the purified substance was completely destroyed by treatment of protease, proteinase K, and pronase E, indicating that it is proteinaceous.

Morphological and genetic characterization and the nationwide distribution of the phototrophic dinoflagellate Scrippsiella lachrymosa in the Korean waters

  • Lee, Sung Yeon;Jeong, Hae Jin;You, Ji Hyun;Kim, So Jin
    • ALGAE
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    • v.33 no.1
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    • pp.21-35
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    • 2018
  • The phototrophic dinoflagellate genus Scrippsiella is known to have a worldwide distribution. Here, we report for the first time, the occurrence of Scrippsiella lachrymosa in Korean waters. Unlike the other stains of S. lachrymosa whose cultures had been established from cysts in the sediments, the clonal culture of the Korean strain of S. lachrymosa was established from motile cells. When the sulcal plates of S. lachrymosa, which have not been fully described to date, were carefully examined using scanning electron microscopy, the Korean strain of S. lachrymosa clearly exhibited the anterior sulcal plate (s.a.), right sulcal plate (s.d.), left sulcal plate (s.s.), median sulcal plate (s.m.), and posterior sulcal plate (s.p.). When properly aligned, the large subunit (LSU) rDNA sequence of the Korean strain of S. lachrymosa was ca. 1% different from those of two Norwegian strains of S. lachrymosa, the only strains for which LSU sequences have been reported. The internal transcribed spacer (ITS) rDNA sequence of the Korean strain of S. lachrymosa was also ca. 1% different from those of the Scottish and Chinese strains and 3% different from those of the Canadian, German, Greek, and Portuguese strains. Thus, the Korean S. lachrymosa strain has unique LSU and ITS sequences. The abundances of S. lachrymosa in the waters of 28 stations, located in the East, West, and South Sea of Korea, were quantified in four seasons from January 2016 to October 2017, using quantitative real-time polymerase chain reaction method and newly designed specific primer-probe sets. Its abundances were >$0.1cells\;mL^{-1}$ at eight stations in January and March 2016 and March 2017, and its highest abundance in Korean waters was $26cells\;mL^{-1}$. Thus, S. lachrymosa has a nationwide distribution in Korean waters as motile cells.

Molecular Phylogeny of the Amynthas-complex (Oligochaeta: Megascolecidae) Inferred from ITS Nucleotide Sequences (Ribosomal DNA ITS 유전자를 이용한 왕지렁이(빈모강: 지렁이과) 그룹의 계통분류)

  • Hong, Yong;James, Samuel W.;Hwang, Ui-Wook;Lee, Bo-Eun;Park, Soon-Cheol;Kim, Tae-Heung
    • Korean Journal of Environmental Biology
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    • v.25 no.4
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    • pp.349-355
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    • 2007
  • Phylogeny of the species mainly from the genus Amynthas in family Megascolecidae was inferred at the molecular level using ITS regions in rDNA. With 26 species of earthworms from 10 genera in 2 families, a stretch comprising the 3'-end of the 18S rRNA, ITS1, 5.8S rRNA, ITS2, and 5' end of 28S rRNA was amplified by applying the primers ITS-1, ITS-2. Phylogenetic analyses of nucleotide sequences with a help of MP, NJ, and QP yielded 5 groups similarly. Genus Amynthas was separated largely into two groups, Korean and Philippine origins. Species grouped into the 1st were Amynthas jirensis, A. agrestis, A. gucheonensis, A. sopaikensis, A. bubonis, A. multimaculatus, A. koreanus, A. dageletensis, A. heteropodus, A. odaesanensis, Pontoscolex sp., Pheretima sp. 1, and Dendropheretima banahawensis. Amynthas halconensis, A. isarogensis, A. mindrooensis, Pithemera sp. 2, Pithmera sp. 1, and Pleionogaster sp. clustered into one clade forming the 2nd group. Polypheretima sp. 1 and polypheretima. sp. 2 stayed closely together representing a separate monophyletic status, forming the 3rd group, apart from species in other genera. Archipheretima sp. falls into the 4th group. Distinct morphological characteristics from Archipheretima also coinsides with its branching away from others in the previously reported molecular analyses. Similar to Perionyx excavatus that has been selected as an outgroup, Aporrectodea tuberculata also showed a long branch in the phylogram, but it differed from other 24 species included in the analyses. Unlike others, for example, its habitat is very closely related to that of man.

Detection of Mycobacterium avium ssp paratuberculosis in Korean Cattle by the Polymerase Chain Reaction (한우 혈액에서 PCR을 이용한 Mycobacterium avium ssp paratuberculosis의 검출)

  • Kim, Kwang-Hyun;Kwak, Kil-Han;Song, Hee-Jong;Cho, Jeong-Gon
    • Journal of Veterinary Clinics
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    • v.27 no.1
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    • pp.23-28
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    • 2010
  • Mycobacterium avium ssp paratuberculosis, intracellular bacteria that can cause chronic granulomatous enteritis in cattle, continues to pose significant economic losses and health problem with high prevalence. The purpose of this study is the polymerase chain reaction (PCR)-base strategy for early detection of M. avium ssp paratuberculosis in whole blood. Blood samples were collected from korean cattles in Jeonbuk, Korea. The 16 out of 88 serum samples were detected M. partuberculosis by ELISA. Then samples of infected 8 Korean cattles were amplified by PCR. The PCR amplified targets are 16s rDNA and heat shock protein 65kDa (hsp 65). The 16s rDNA provided a highly sensitive and specific tool for the direct detection of mycobacteria. In addition M. avium was confirmed characteristically by the hsp65. Finally there were sure to M. avium ssp paratuberculosis by IS900 PCR. The restriction fragment length polymorphism was identified by PCR amplifications and subsequence restriction enzyme digestions with Pst I of a hsp65. These results indicate that confirm M. avium with 16s rDNA, hsp65 and a restriction fragment length polymorphism in the hsp65 gene can be seem the other pattern. Therefore, these results can be used for clinical direct detections of M. avium ssp paratuberculosis in whole blood of Korean cattle and also to be used epidemiological researches.

The Determination of the Partial 28S Ribosomal DNA Sequences and Rapid Detection of Phellinus linteus and Related species

  • Park, Hyung-Sik;Kim, Gi-Young;Nam, Byung-Hyouk;Lee, Sang-Joon;Lee, Jae-Dong
    • Mycobiology
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    • v.30 no.2
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    • pp.82-87
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    • 2002
  • Species of Phellinus were known to harmful fungi causing white pocket rot and severe plant disease such as canker or heartrot in living trees in the West, but some species have been used to traditional medicines in the Orient for a long time. In this study the partial D1-D2 nucleotide sequences of 28S ribosomal DNA from 13 Phellinus strains were determined and compared with the sequences of 21 strains obtained from GenBank database. According to the neighbor-joining(NJ) method comparing the sequence data the phylogenetic tree was constructed. The phylogenetic tree displayed the presence of four groups. Group I includes P. ferreus, P. gilvus and P. johnsonianus, Group II contains P. laevigatus, P. conchatus and P. tremulae, Group III possesses P. linteus, P. weirianus, P. baumii, P. rhabarbarinus and P. igniarius, and Group IV comprises P. pini, P. chrysoloma. P. linteus and P. baumii, which were used mainly in traditional medicine, belong to the same group, but exactly speaking both were split into two different subgroups. To detect P. linteus only, we developed the PCR primer, D12HR. The primer showed the specific amplification of P linteus, which is permitted to medicinal mushroom in the East. The results make a potential to be incorporated in a PCR identification system that could be used for the rapid identification of this species from its related species, P. linteus especially.

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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    • v.36 no.2
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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.

Pucciniastrum hydrangeae-petiolaris, a Newly Found Rust Fungus on Hydrangea petiolaris in Korea (국내 미기록 등수국 녹병균 Pucciniastrum hydrangeae-petiolaris)

  • Lee, Jae Sung;Choi, Young-Joon;Choi, Byoung-Ki;Jung, Bok-Nam;Park, Ji-Hyun;Shin, Hyeon-Dong
    • The Korean Journal of Mycology
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    • v.49 no.1
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    • pp.119-125
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    • 2021
  • Hydrangea petiolaris, belonging to the family Hydrangeaceae, is a vine plant distributed in Ulleung, Jeju, and other southern islands of Korea. In October 2017, a rust fungus was discovered on H. petiolaris in Jeju Island, Korea. To identify the rust fungus, we performed a morphological examination and molecular phylogenetic analysis of the internal transcribed spacer and 28S large subunit rDNA sequences. As a result, the fungus was identified as Pucciniastrum hydrangeae-petiolaris, consistent with previous reports from Japan and Russia, but showed a significant phylogenetic distance from Pucciniastrum hydrangeae reported on Hydrangeae spp. in North America. This is the first record of P. hydrangeae-petiolaris on H. petiolaris in Korea.

Molecular Characterization of the Bacterial Community in Activated Sludges by PCR­RFLP (PCR-RFLP 방법을 이용한 활성 슬러지의 세균군집 분석)

  • Lee Hyun-Kyung;Kim Jun-Ho;Kim Chi-Kyung;Lee Dong-Hun
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.307-312
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    • 2004
  • Diversity of the bacterial communities and the relation between community structure and components of waste­water were analyzed by 16S rRNA-based molecular techniques. Clone libraries of the 16S rDNAs from the sludges were constructed by PCR and cloning. The 1,151 clones from a sludge sample of sewage treatment plant were clustered into 699 RFLP phylotypes and the 1,228 clones from the wastewater disposal plant of chemical industry were clustered into 300 RFLP phylotypes. Shannon-Weiner diversity indices of two sampling sites were 8.7 and 6.1, indicating that the bacterial community structure of sewage treatment plant was more diverse than that of wastewater disposal plant of chemical industry. Forty clones belonging to predominant RFLP types were selected and sequenced. Seventy percent (28 clones) of the sequenced clones were related to the uncultured bacteria in public databases. The ${\beta}-Proteobacteria$ dominated in the bacterial communities of investigated two sludge samples. 16S rDNA sequences of the sewage treatment plant were similar to those of other activated sludges, while the bacterial community in wastewater disposal plant of chemical industry rep­resented the strains identified from high-temperature, anaerobic, hydrocarbon-rich, and sulfur-rich environ­ments. This result suggested that bacterial communities depended upon the components of wastewater.

Molecular Biological Diagnosis of Meloidogyne Species Occurring in Korea

  • Oh, Hyung-Keun;Bae, Chang-Hwan;Kim, Man-Il;Wan, Xinlong;Oh, Seung-Han;Han, Yeon-Soo;Lee, Hyang-Burm;Kim, Ik-Soo
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.247-255
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    • 2009
  • Root-knot nematode species, such as Meloidogyne hapla, M. incognita, M. arenaria, and M. javanica are the most economically notorious nematode pests, causing serious damage to a variety of crops throughout the world. In this study, DNA sequence analyses were performed on the D3 expansion segment of the 28S gene in the ribosomal DNA in an effort to characterize genetic variations in the three Meloidogyne species obtained from Korea and four species from the United States. Further, PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism), SCAR (Sequence Characterized Amplified Region) PCR and RAPD (Randomly Amplified Polymorphic DNA) were also utilized to develop methods for the accurate and rapid species identification of the root-knot nematode species. In the sequence analysis of the D3 expansion segment, only a few nucleotide sequence variations were detected among M. incognita, M. arenaria, and M, javanica, but not M. hapla. As a result of our haplotype analysis, haplotype 5 was shown to be common in M. arenaria, M. incognita, M. javanica, but not in the facultatively parthenogenetic species, M. hapla. PCR-RFLP analysis involving the amplification of the mitochondrial COII and large ribosomal RNA (lrRNA) regions yielded one distinct amplicon for M. hapla at 500 bp, thereby enabling us to distinguish M. hapla from M. incognita, M. arenaria, and M. javanica reproduced via obligate mitotic parthenogenesis. SCAR markers were used to successfully identify the four tested root-knot nematode species. Furthermore, newly attempted RAPD primers for some available root-knot nematodes also provided some species-specific amplification patterns that could also be used to distinguish among root-knot nematode species for quarantine purposes.