• Title/Summary/Keyword: 16S ribosomal DNA

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Candidatus Phytoplasma trifolii Associated with Witches' broom of Lespedeza cyrtobotrya M.

  • Kim, Young-Hwan;Jung, Hee-Young
    • The Plant Pathology Journal
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    • v.23 no.2
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    • pp.106-108
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    • 2007
  • The Symptoms of witches' broom disease caused by phytoplasma including general stunting and yellowing, were observed in leafy lespedeza (Lespedeza cyrtobotrya M.) on Doam-myeon, Pyeongchang-gun, in 2006. Based on the sequence analysis of PCR-amplified 16S ribosomal DNA and 16S-23S spacer region DNA products using universal phytoplasma primers, the phytoplasma associated with leafy lespedeza witches' broom (LLWB) disease was identified as a member of Candidatus Pytoplasma trifolii. It was most closely related to alsike clover proliferation phytoplasma (99.8% similarity, accession no. AY390261), Candidatus Pytoplasma trifolii strain. RFLP patterns generated with AluI, HpaII clearly differentiated LLWB phytoplasma from the referenced phytoplasma strains, water dropwort witches' broom, mulberry dwarf, glehni aster yellow dwarf and jujube witches' broom. This paper is the first report on Candidatus Phytoplasma trifolii in leafy lespedeza identified at a molecular level.

Development of Species-specific PCR Primers for Detecting Peptoniphilus mikwangii

  • Park, Soon-Nang;Lee, Junhyeok;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.42 no.3
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    • pp.143-147
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    • 2017
  • In a previous study, Peptoniphilus mikwangii was isolated from the human oral cavity as a new species. The purpose of this study was to develop P. mikwangii-specific PCR primers. The PCR primers were designed, based on the nucleotide sequence of 16S ribosomal RNA (16S rDNA). The specificity of the primers was tested using genomic DNAs of 3 strains of P. mikwangii and 27 strains (27 species) of non-P. mikwangii bacteria. The sensitivity of primers sensitivity was determined using PCR, with serial dilutions of the purified genomic DNAs (4 ng to 4 fg) of P. mikwangii KCOM $1628^T$. The data showed that P. mikwangii-specific qPCR primers (B134-F11/B134-R1 & B134-F5/B134-R5) could detect only P. mikwangii strains, and 400 fg or 40 fg of P. mikwangii genome DNA. These results suggest that PCR primers are useful in detecting P. mikwangii from the oral cavity.

Practical considerations for the study of the oral microbiome

  • Yu, Yeuni;Lee, Seo-young;Na, Hee Sam
    • International Journal of Oral Biology
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    • v.45 no.3
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    • pp.77-83
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    • 2020
  • In the oral cavity, complex microbial community is shaped by various host and environmental factors. Extensive literature describing the oral microbiome in the context of oral health and disease is available. Advances in DNA sequencing technologies and data analysis have drastically improved the analysis of the oral microbiome. For microbiome study, bacterial 16S ribosomal RNA gene amplification and sequencing is often employed owing to the cost-effective and fast nature of the method. In this review, practical considerations for performing a microbiome study, including experimental design, molecular analysis technology, and general data analysis, will be discussed.

Vibrio 4종의 16s-23s ribosomal DNA Intergenic Spacer region의 유전학적 분석

  • 장인권;이실한;이승환;최용석;임현정;송재희
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.163-164
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    • 2000
  • Vibrio는 새우양식장 및 종묘배양장의 사육수에서 가장 qlsqjsgkrp 출현하며 양식새우의 질병과 직간접적으로 관련을 갖고 있는 주요 병원성 세균류로서 이들 종의 신속한 동정은 질병의 조기진단 및 대책을 위하여 매우 중요하기 때문에 이에 대한 연구가 많이 이루어져 왔으나 일부 Vibrio 종들간의 생리, 화학적 특성이 너무도 유사하여 동정에 많은 어려움을 겪고 있다. (중략)

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Morphological and Molecular Classifications of Genus Pholis

  • Lee, Sung-Hoon;Jang, Yo-Soon;Baik, Chung-Boo;Han, Kyeong-Ho;Myung, Jung-Goo;Lee, Jin-Hee;Choi, Sang-Duk;Kim, Seon-Jae;Kim, Jong-Oh;Hwang, Jae-Ho
    • Animal cells and systems
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    • v.13 no.4
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    • pp.453-460
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    • 2009
  • Morphological and molecular classifications were attempted in an effort to establish species-specific classifications of three species of the genus Pholis in Korea; these species were subjected to morphological and molecular methodologies using body measurements, RFLP, RAPD, and phylogenetic trees using the nucleotide sequences of mitochondrial 16S and 12S ribosomal DNAs, cytochrome c oxidase I, and cytochrome b. The data demonstrated that the three species of genus Pholis are distinct from each other, both morphologically and genetically.

Monitoring of Commercial Cephalopod Products Sold on the South Korea Market using DNA Barcode Information (DNA 바코드를 이용한 국내 유통 두족류 제품의 원재료 모니터링 연구)

  • Yu, Yeon-Cheol;Hong, Yewon;Kim, Jung Ju;Kim, Hyung Soo;Kang, Tae Sun
    • Journal of Food Hygiene and Safety
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    • v.34 no.5
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    • pp.502-507
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    • 2019
  • Cephalopods are one of the most important fishery resources in the world because of their desirable taste and nutritional value. In south Korea, one of the countries in which a large amount of seafood is consumed, cephalopods (e.g., octopus, squid, and cuttlefish) have an annual consumption rate of over 400,000 metric tons. In this study, octopus and squid products (n=28) sold on the market were monitored by analyzing sequences of DNA barcode markers (cytochrome c oxidase subunit I and 16S ribosomal RNA genes). For species identification, the NCBI BLAST database was screened with the sequences and analyzed as a query. In this BLAST search, twelve squid products showed 99-100% sequence identity to Dosidicus gigas (n=3) and Todarodes pacificus (n=9). In the case of the other 16 products that were declared using octopus as raw materials on the labels, six products were identified as Cistopus taiwanicus (n=1), Amphioctopus marginatus (n=1), Scaeurgus unicirrhus (n=1), and Dosidicus gigas (n=3). Monitoring results indicated that a significant percentage (37.5%) of mislabeling was present in octopus products sold on the South Korean market.

Genetic Variation in Mutants Induced by Gamma Ray in Hypsizigus marmoreus (느티만가닥버섯에서 감마선에 의한 돌연변이체들의 유전적 변이)

  • Kim, Jong-Bong;Yu, Dong-Won
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1174-1179
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    • 2014
  • This research was carried out to evaluate whether gamma ray is a useful tool for breeding new strains of mushrooms. For this research, 5 mutant groups, 20 strains of Hypsizigus marmoreus, 2 strains of Lyophyllum decastes, and 1 strain of Lyophyllum shimeji were used. Monokaryon spores from one variety of H. marmoreus were irradiated with 50~2,000 Gy of gamma ray. The propriety dose was 50~200 Gy for mutagenesis. Mutant monokaryon mycelia crossed each order to become dikaryon mycelia. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR, and the products were sequenced. The sequences of the ITS regions (16 partial rDNA, complete ITS1, 5.8 rDNA and partial rDNA) were analyzed by PCR, and strains of H. marmoreus, L. decastes, and L. shimeji were auto-sequenced. The lengths of the sequenced ITSs were 1,052~1,143 nucleotides. Genetic matrices were calculated using Nei-Li's genetic distance coefficient based on ITS sequence. The dissimilarities were 0~3.35% in strains of H. Hypsizigus. In addition, a phylogenetic tree was constructed based on ITS sequences using the neighbor-joining (NJ) method. The phylogenetic tree revealed that 23 strains and 5 mutant groups were divided into 12 clusters; the mutant groups fell into different clusters. These results show that mushroom spores were mutated effectively by gamma ray; therefore, gamma ray could be a useful tool for breeding new strains of mushrooms.

Nested PCR for the Detection of Streptococcus mutans (Nested PCR를 이용한 Streptococcus mutans의 검출)

  • Choi, Min-Ho;Yoo, So-Young;Lim, Chae-Kwang;Kang, Dong-Wan;Kook, Joong-Ki
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.19-25
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    • 2006
  • This study was undertaken to develop PCR primers for the identification and detection of Streptococcus mutans (by)using species-specific forward and universal reverse primers. These primers targeted the variable regions of the 16S ribosomal RNA coding gene (rDNA). The primer specificity was tested against 11S. mutans strains and 10 different species (22 strains) of oral bacteria. The primer sensitivity was determined by testing serial dilutions of the purified genomic DNA of S. mutans ATCC $25175^T$. The data showed that species-specific amplicons were obtained from all the S. mutans strains tested, which was not observed in the other species. The direct and nested PCR could detect as little as 2 pg and 2 fg of the chromosomal DNA from S. mutans ATCC $25175^T$, respectively. This shows that the PCR primers are highly sensitive and applicable to the detection and identification of S. mutans.

Seasonal Differences of Bacterial Communities Associated with the Marine Sponge, Hymeniacidon sinapium (주황해변해면(Hymeniacidon sinapium) 공생세균 군집의 계절적 차이)

  • Jeong, Jong-Bin;Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.48 no.4
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    • pp.262-269
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    • 2012
  • Seasonal differences of the cultivable bacterial communities associated with the marine sponge, Hymeniacidon sinapium, between spring and summer were analyzed through the Amplified Ribosomal DNA Restriction Analysis (ARDRA). For the cultivation of the bacterial isolates, modified Zobell and MA media were used. The 16S rDNA of individual strains were amplified and fragmented by using two restriction enzymes, HaeIII and MspI. As a result, 23 ARDRA types from the spring sponge and 28 types from the summer sponge were obtained. The partial sequencing result of 1 to 3 selected strains from each types showed over 94% similarities with the known species from the public database. The bacterial communities from the sponge, captured on spring, contained 4 phyla: Actinobacteria, Alphaproteobacteria, Gammaproteobacteria, and Firmicutes. There were 5 phyla observed from the bacterial communities associated with the sponge, captured on summer: Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Bacteroidetes. Gammaproteobacteria was predominant group in both spring and summer, accounted for 33.8% of total in spring and 67.4% in summer, showed increase pattern on summer. Because Firmicutes and Actinobacteria participated in 30.2% and 8.3% of the spring sponge while they represented only 6.9% and 0% of the summer sponge, both bacterial groups showed decrease drift on summer. Betaproteobacteria (4.7%) and Bacteroidetes (4.7%) were only observed on the sponge captured on summer. On the sponge, Hymeniacidon sinapium, more diverse bacterial communities were shown on summer than on spring, and even from the same sponge, there were seasonal differences.

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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    • v.34 no.2
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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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