• Title/Summary/Keyword: l6S rDNA

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IDENTIFICATION OF PUTATIVE PATHOGENS IN ACUTE ENDODONTIC INFECTIONS BY PCR BASED ON 16S rDNA (중합효소연쇄반응법을 이용한 급성 치수 및 치근단 질환의 병원성 세균의 동정)

  • Kim, Ji-Hoon;Yoo, So-Young;Lim, Sun-A;Kook, Joong-Ki;Lim, Sang-Soo;Park, Seul-Hee;Hwang, Ho-Keel
    • Restorative Dentistry and Endodontics
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    • v.28 no.2
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    • pp.178-183
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    • 2003
  • The purpose of this study was to investigate the frequency of 7 putative pathogens in endodontic infections. The specimens were collected from infected pulpal tissue of patients who were referred for root canal treatment to the department of conservative dentistry, Chosun University Samples were collected aseptically using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ml of 1 X PBS. DNAs were extracted from the samples by direct DNA extraction method using lysis buffer (0.5% EDTA, 1% Triton X-100). Identification of 7 putative pathogens was performed by PCR based on 16S rDNA. The target species were as follows : Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, and Treponema denticola. Our data revealed that the prevalence of P. endodontalis was found in 88.6% (39/54), P. ginivalis 52.3% (23/44), P. nigrescens 18.2% (8/44), P intermedia 15.9% (7/44) B. forsythus 18.2% (8/44), A. actinomycetemcomitans 3.3% (1/44), T. denticola 25% (l1/44) of the samples. The high prevalence of P. endodontalis and P. ginivalis suggests that they may play an important role in the etiology of endodontic infections.

Taxonomic status of Goodyera rosulacea (Orchidaceae): molecular evidence based on ITS and trnL sequences (로젯사철란(Goodyera rosulacea: Orchidaceae)의 분류학적 위치: ITS와 trnL 염기서열에 의한 분자적 증거)

  • Lee, Chang Shook;Eom, Sang Mi;Lee, Nam Sook
    • Korean Journal of Plant Taxonomy
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    • v.36 no.3
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    • pp.189-207
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    • 2006
  • Goodyera rosulacea, which is morphologically similar to G. repens, is described recently as a new species based on its distinct morphological characters such as rosette-formed leaves, short rhizome and habitat. To verify the taxonomic identity of G. rosulacea and its taxonomic relationship within Korean Goodyera taxa, sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA and the trnL region of cpDNA from 24 accessions including 1 outgroup accession were analyzed. Aligned sequences were analyzed using maximum parsimony and distance method, and the taxonomic identity and the taxonomic relationships among the related taxa were estimated by the existence of private marker gene and the phylogenetic tree of the aligned sequences. Molecular data indicate that G. rosulacea gas several private marker genes and shows monophyly in phylogenetic trees of both ITS and trnL sequences. the pairwise distance between G. rosulacea and the orher taxa of Korean Goodyera was 3.49-6.68% for ITS region and 5.05-9.53% for trnL region, indicating that G. rosulacea could be treated as an independent species. Therefore, our molecular data support the taxonomic of G. rosulacea as a distinct species of Korea. In phylogenetic trees, G. rosulacea formed same clade with G. repens, which has similar morphological characters with G. rosulacea, and showed the lowest pairwise distance with G. repens among Korean Goodyera taxa. These molecular data sugguested that G. rosulacea and G. repens are closely related taxa.

Classification and Characterization of Bacteriophages of Lactobacillus casei -Analysis of Restriction Patterns of Phage DNA- (Lactobacillus casei bacteriophage의 분류 및 특성에 관한 연구 - Phage DNA의 제한효소 절편 비교 분석-)

  • 김영창;강현삼
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.115-121
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    • 1985
  • Five representative virulent phages (J1, TK93, K1, PD5, and CP1) and one temperate phage (.phi.1043) of Lactobacillus casei were compared to each other by analyzing the agarose gel electrophoretic patterns of restriction enzyme-digested phage DNAs. Nucleic acids of all the tested phages were double stranded DNA. DNAs of J1, TK93, K1, and ${\phi}$ 1043 phages had a size of about 42kb, but the size of PD5 and CP1 DNAs was avout 140kb. J1, TK93, K1, PD5, CP1, and ${\phi}$ 1043 DNAs were digested to 13, 13, 11, 14, 14, and 12 fragments by EcoR1, respectively, and showed its characteristec restriction patterns. Cohesive ends were present in J1, TK93, and ${\phi}$ 1043, but were absent in K1, PD5, and CP1. Restriction maps of J1 and TK93 DNAs showed nearly complete homology and their evolutionary relationship based upon the restriction analysis was discussed.

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Physical Characteristics and Antagonistic Effect of Ampelomyces (Ampelomyces의 생리적 특성 및 길항능력)

  • Kim, Ji-Young;Lee, Wang-Hyu;Kim, Hyung-Moo
    • Research in Plant Disease
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    • v.15 no.3
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    • pp.209-216
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    • 2009
  • During the period of June, 2005 to May, 2008, 44 host plants infected with powdery mildew were collected in the Jeon-ju and Jang-su districts of Jeonbuk province and in the Jang-sung district of Jeonnam province. The hyperparasites, Ampelomyces were confirmed in 12 plant species. Most of the pycnidium shapes of Ampelomyces were circular or oval shaped, and the sizes were different even within the same host plant, and also the color of pycnidium was ranged from light brown to dark brown. Ampelomyces species were isolated from 4 hosts including Impatiens balsamina L., Cucurbita pepo, Rudbeckia laciniata var. elatier and Youngia sonchifolia, and thus the most appropriate 12 Ampelomyces strains for the current experiment were selected. When analyzing the selected 12 strains' incubational and nutritional characteristics, the malt extract agar was the most appropriate media. When investigating the effect of osmotic pressure on the spore germination, 0.15M NaCl concentration was the optimum germination concentration. When the isolated Ampelomyces sp. was tested in-vitro, it was found to be effective to control in other plant pathogens, isolated Ampelomyces showed no pathogenicity to the plant. strains isolated . studied on rDNA ITS sequence analysis. The rDNA ITS sequence data of Ampelomyces sp. isolate BSLAH16 from Impatiens balsamina L. were analyzed and identified.

A case of parentage testing in the Thoroughbred horse by microsatellite DNA typing (Microsatellite DNA형에 의한 더러브렛 말의 친자감정예)

  • Cho, Gil-Jae;Yang, Young-Jin;Kim, Bong-Hwan
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.25-29
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    • 2003
  • This study was carried out to investigate a usefulness of the microsatellite DNA markers for parentage verification of Thoroughbred (TB) horses. 9 TB horses samples were genotyped for nine international minimum standard markers (AHT4, 5, ASB2, HMS3, 6, 7, HTG4, 10, and VHL20), and the additional panel of four markers, ASB17, CA425, LEX33, and TKY321. This methods consisted of multiplexing PCR procedures, and it showed reasonable amplification of all PCR products. Genotyping was performed with an ABI 310 genetic analyzer. Foal I was excluded according to principles of Mendelian genetics in AHT4 (H/K), ASB2 (Q/Q), HMS3 (I/P), HTG4 (M/O), HTG1O (K/R), VHL20 (M/P), ASB17 (F/N), LEX33 (M/O), and TKY321 (G/I) markets. Foal II was excluded with markers AHT5 (K/M), ASB2 (M/N), HMS7 (N/N), HTG1O (K/K), VHL20 (I/I), ASB17 (F/F) and TKY321 (G/I). Foal III was excluded with markers AHT4 (O/O), AHT5 (K/K), ASB2 (M/R), HMS6 (M/P), HMS7 (O/O), HTG10 (R/S), VHL20 (L/M), and ASB17 (N/O). These results suggest that the present DNA typing is so useful for parentage verification of TB horses.

Isolation and Characterization of Aeromons hydrophila PBl6 and Properties of Synthetic Wastewater Degradation (Protease 생성균 Aeromonas hydrophila PB16의 분리 및 합성폐수처리능)

  • 박형수;양선영;김무훈;이종광;유용호;박두현
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.235-240
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    • 2002
  • Protease producing bacterium, PB16 was isolated from food processing wastewater sludge and paddy field soil samples and selected by the clear zone and enzyme activity test. The isolate was gram negative, rod type and its protease productivity was 6.49 U/ml. As a result of API20NE kit test and 16S rDNA sequencying, the isolated PB16 was identified as Aeromonas hydrophila (99%). The growth rate ($h^{-1}$) was 0.21 in synthetic waste water only and 0.26 in synthetic waste water containing vitamin and mineral using a bioscreen C. Synthetic wastewater removal rate was 59 and 87%, respectively after 1 and 3 day reaction (intial CODcr was 2,472 mg/l).

A Commensal Thermophile, Symbiobacterium toebii: Distribution, Characterization, and Genome Analysis

  • Bae Jin-Woo;Kim Kwang;Song Jae Jun;Ha Jae Seok;Kim Joong-Jae;Kang Gwan-Tae;Kim Mi-Hwa;Hong Seung-Pyo;Sung Moon-Hee
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2001.11a
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    • pp.46-53
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    • 2001
  • A commensal thermophile, Symbiobacterium toebii, isolated from hay compost (toebii) in Korea commensally interacted with a thermophilic Geobacillus toebii sp. nov., which was a new species within the genus Geobacillus on the basis of the phenotypic traits and molecular systematic data. S. toebii required the crude extracts and/or culture supernatant of the Geobacillus toebii for axenic growth and could grow on the temperature between 45 and $70^{\circ}C$ (optimum: $60^{\circ}C$; 2.4 h doubling time) and pH 6.0 and 9.0 (optimum: pH 7.5). The G+C content of the genomic DNA was $65 mol\%$, and the major quinones were MK-6 and MK-7. A phylogenetic analysis of its 16S rDNA sequence indicated that Symbiobacterium toebii was closely related with solely reported Symbiobacterium thermophilum. The presence of the commensal thermophile 16S rDNA and accumulation of indole in all the enriched cultures indicate that Symbiobacterium toebii is widely distributed in the various soils. The genome of S. toebii constituted a circular chromosome of 3,280,275 base pairs and there was not an extra-chromosomal element (ECE). It contained about 4,107 predicted coding sequences. Of these protein coding genes, about $45.6\%$ was encoded well-known proteins and annotated the functional assignment of 1,874 open reading frames (ORFs), and the rest predicted to have unknown functions. The genes encoding thermostable tyrosine phenol-lyase and tryptophan indole-lyase were cloned from the genomic DNA of S. toebii and the enzymatic production of L-tyrosine and L-tryptophan was carried out with two thermostable enzymes overexpressed in recombinant E. coli.

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Characterization of a Rhizobacterium Promoting Early Growth in Maize (옥수수 초기 성장을 촉진하는 근류세균의 특성)

  • Lee Sang-Eun;Yi Hwe-Su;Park Seung-Hwan;Ghim Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.70-73
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    • 2005
  • A soil bacterium was isolated from maize roots cultivated in Korea (KNUC153). The isolate was partially classified on basis of l6S rDNA sequence analysis as Stenotrophomonas maltophilia. By the acetylene reduction assay (ARA), the strain KNUC153 contained nitrogen-fixing abilities. The amount of auxin produced by the strain KNUC153 was $77.6\;{\mu}g/ml$. The strain KNUC153 produced 4 times higher amount of l-amino-cyclopropane-l-carboxy­lic acid deaminase than that of the other known strain Azospirillum sp. KNUC82. Inoculation treatment with the strain KNUC153 for maize seeds showed positive effect on early growth of the plants.

Biodegradation of Diesel with Pseudomonas sp, KDi19 in Liquid Medium (Pseudomonas sp. KDi19를 이용한 액체배지내에서 경유의 생물학적 분해)

  • Yun, Min-Woo;Jeong, Jeong-Hwa;Chang, Soon-Woong;Kong, Sung-Ho;Lee, Jong-Yeol;Kang, Dong-Hyo;Lee, Sang-Seob
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.12
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    • pp.1285-1291
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    • 2005
  • In this study, we isolated bacteria from petroleum contaminated soil which were near to underground storage tanks(UST). Through the screen test, we selected high efficiency bacterium, KDi19, for biodegradation of diesel. KDi19 was identified as Pseudomonas sp. by 16S rDNA, fatty acid, and morphological physiological characteristics. KDi19 degraded 956.3 mg/L(95.6%) of 1,000 mg/L diesel for 48 hours(incubation condition : temperature; $30^{\circ}C$, cell concentration; 1.0 g/L, pH 7). At low temperature, $20^{\circ}C$, $15^{\circ}C$, $10^{\circ}C$, KDi19 respectively removed 63.9%, 18.5% and 17.0% of 1,000 mg/L diesel for 48 hours(cell concentration 1.0 g/L, pH 7). At low concentration of diesel, 50 mg/L and 100 mg/L, KDi19 degraded 97.9% and 96.2% of diesel for 24 hours(temperature; $30^{\circ}C$, cell concentration: 1.0 g/L, pH 7), respectively.

Isolation and Characterization of Lactobacillus brevis AML15 Producing γ-Aminobutyric acid ((γ-Aminobutyric acid를 생산하는 Lactobacillus brevis AML15의 분리 및 특성)

  • Shin, Ji-Won;Kim, Dong-Geol;Lee, Yong-Woo;Lee, Hyoung-Seok;Shin, Kee-Sun;Choi, Chung-Sig;Kwon, Gi-Seok
    • Journal of Life Science
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    • v.17 no.7 s.87
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    • pp.970-975
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    • 2007
  • For the screening of ${\gamma}-aminobutyric$ acid (CABA)-producing bacteria, 86 bacterial strains which produce GABA were isolated from Kimchi and Salted fisk .Among these, three strains designated AML15, AML45-1, AML72 with relatively high GABA productivity were selecled by thin layer chromatography (TLC). To elucidate the relationship between isolated strains and the genus Lactobacillus, their 16S rDNA sequence were examined. The result of their DNA sequences showed 99% similarity with Lactobacillus brevis ATCC 367. On the basis of the these results, isolated strains were identified as Lactobacillus brevis and designated L. brevis AML15. In order to determine the optimum conditions for GABA production, the isolated strains were cultivated in pyridoxal phosphate (PLP) and monosodium glutami. acid (MSG). Results showed that L. brevis AML15 had the highest CABA productivity with 10,424 $nM/{\mu}l$ concentration in MRS broth containing 5% (w/v) MSG and 10 ${\mu}M$ PLP at pH 5.0. The results imply that L. brevis AML15 has the potential to be developed as a strain for GABA hyper-production.