• Title/Summary/Keyword: r-DNA

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Community Structure of Bacteria Associated with Two Marine Sponges from Jeju Island Based on 16S rDNA-DGGE Profiles (16S rDNA-DGGE를 이용한 2종의 제주도 해양 해면의 공생세균의 군집 구조)

  • Park, Jin-Sook;Sim, Chung-Ja;An, Kwang-Deuk
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.170-176
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    • 2009
  • Culture-independent 16S rDNA-DGGE profiling and phylogenetic analysis were used to examine the predominant bacterial communities associated with the two sponges, Dictyonella sp. and Spirastrella abata from Jeju island. The culture-independent approach involved extraction of total bacterial DNA, PCR amplification of the 16S ribosomal DNA using primer pair 341f-GC and 518r, and separation of the amplicons on a denaturing gradient gel. Denaturing gradient gel electrophoresis banding patterns indicated 8 and 7 bands from the two sponge species, Dictyonella sp. and Spirastrella abata, respectively. There were not common major bands in two different sponges. Comparative sequence analysis of variable DGGE bands revealed from 93% to 98% similarity to the known published sequences. The dominant bacterial group of Dictyonella sp. belonged to uncultured Gammaproteobacteria, while, that of Spirastrella abata belonged to uncultured Alphaproeobacteria and Firmicutes. DGGE analysis indicated predominant communities of the sponge-associated bacteria differ in the two sponges from the same geographical location. This result revealed that bacterial community profiles of the sponges were host species-specific.

Molecular Biological Identification of Bacteria in Middle Ear Effusion Using 16S rDNA Multiplex PCR (중이 삼출액 미생물의 16S rDNA 복합중합효소연쇄반응을 이용한 분자생물학적인 진단)

  • 이정구;이인숙;박지연;정상운;오충훈
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.36-39
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    • 2003
  • The rapid and reliable 16S rDNA multiplex polymerase chain reaction (PCR) assay was established to characterize bacterial etiologies of middle ear effusion. These etiologies included Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumonia, which were detected in middle-ear effusion (MEE) samples taken from patient with otitis media. A total of 39 MEE samples were aspirated from 26 patients. DNA was extracted from MEE samples, and PCR was done with DNA extracts by using the common primers, which is localized at C4 region in the 16S rDNA gene of all bacterial species, and species-specific primers: (i) Haemophilus-specific primer, (ii) Moraxella- specific primer, and (iii) Streptococcus-specific primer. Among 39 samples tested, 24 (61.5%) were positive for H. influenzae, 10 (25.6%) were positive for M. catarrhalis, 3(7.7%) were positive for S. pneumonia, and 11 (28%) were negative for 165 rDNA multiplex PCR reaction. Nine samples (28.6%) exhibited a mixed infection and were positive for both H. infuenzae and M. catarrhalis. We suggested that 16S rDNA multiplex PCR is a useful method to identify rapidly for rapid identification of the pathogenic bacteria and characterization of bacterial etiologies of middle ear effusion.

DNA Barcoding of Scolelepis (Parascolelepis) papillosa (Annelida, Spionidae) in Korea, with Additional Taxonomic Notes

  • Lee, Geon Hyeok;Lee, Ha-Eun;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.37 no.4
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    • pp.349-353
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    • 2021
  • Scolelepis (Parascolelepis) papillosa (Okuda, 1937), originally described from a single incomplete individual from Jeju Island in Korea, was collected from the intertidal sandflats of Soan Island (Jeollanam-do province) in Korea. The examined specimens of S. (P.) papillosa agree well with the original description in having the papillae on the basal sheath of the palps, presence of occipital antenna, absence of notochaetae in chaetiger 1, branchiae completely fused with notopodial postchaetal lamellae at the anterior chaetigers, and neuropodial hooded hooks appearing from chaetiger 16. In this study, the sequences of partial mitochondrial cytochrome c oxidase subunit I (COI), 16S ribosomal DNA (16S rDNA), and the nuclear 18S ribosomal DNA (18S rDNA) of the species were determined. We also provide the detailed description and illustrations on this species based on the complete specimens newly collected in this study.

DNA Barcoding of Scolelepis (Scolelepis) sagittaria (Annelida, Spionidae) in Korea, with a Morphological Variability of the Species

  • Lee, Geon Hyeok;Yoon, Seong Myeong;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.38 no.3
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    • pp.144-147
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    • 2022
  • The polychaete Scolelepis (Scolelepis) sagittaria was originally described from Japanese waters and subsequently reported from Korean waters. In this study, we determined for the first time the mitochondrial cytochrome c oxidase subunit I (COI), 16S ribosomal DNA (16S rDNA), and nuclear 18S ribosomal DNA (18S rDNA) sequences of Korean specimens of S. (S.) sagittaria. We also assessed intraspecific variation in the shape of the prostomium of this species based on an examination of 247 individuals. All materials were collected from intertidal sandy beaches of the Korea Strait. The molecular data and morphological observations reported herein will contribute to gaining a better understanding of the taxonomic relationships among members of the genus Scolelepis.

Tracking of the $Km^r$ Gene in Conjugal Transfer by Using DNA Probe (DNA Probe에 의한 $Km^r$ 유전자의 전이 추적)

  • 이성기;김치경
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.483-490
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    • 1992
  • In order to understand the transfer behavior of a particular gene in water environments, kanamycin resistance ($Km^r$) gene was tracked by Southern hybridization with DNA probe in its conjugal transfer. A $Km^r$ natural bacterial isolate and genetically modified microorganisms (GMMs) constructed from the isolate were used as donor for conjugal transfer of the $Km^r$ gene. The transfer frequencies of the $Km^r$ gene from GMM strains were generally 10 to 100 times higher than those from the natural isolate. The conjugants obtained from GMM strains in LB broth had more plasmids newly appeared, and particularly the conjugants in A Wand FW waters revealed more rearrangement in their plasmids as a function of conjugation time. When plasmids of the conjugants obtained in LB broth were Southern hybridized with DNA probe of the $Km^r$ gene, the $Km^r$ plasmids in the conjugants were detected at the same position of the plasmids in donor cells, in spite of the fact that the plasmids were highly rearranged in conjugant cells. But the $Km^r$ plasmids in the donor of DKI and DKC601, and DKC600 were not identified in the conjugants obtained after 50 h conjugation in AW and after 30 h in AW, respectively. The size of the $Km^r$ plasmids showing hybridization signal were a little changed in the conjugants obtained in A Wand FW waters. Therefore, the method of Southern hybridization with DNA probe was proved to be very specific and useful for tracking of particular genes in water environments.

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Identification of Bacteria Causing Fermentation of Oriental Melon in Korea (참외 발효과를 유발하는 세균의 동정)

  • Choi, Jae-Eul;Cha, Sun-Kyung;Kim, Jin-Hee;Yuk, Jin-Ah;Hwang, Yong-Soo;Kwon, Soon-Wo
    • Research in Plant Disease
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    • v.9 no.4
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    • pp.189-195
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    • 2003
  • Bacteria causing fermentation in Oriental melon were identified as three independent groups on the basis of 16S rDNA sequence analysis. The 16S rDNA sequence of the strain CM2105 showed the highest identity (99.6%) with that of Microbacterium phyllosphaerae, and also indicated high sequence identity to that of M. holiorum (99.5%). The 16S rDNA sequences of the strain CM2101 and CM2121 matched at the high sequence similarity (98.9%, 98.8, respectively), to that of Pseudomonas pavonacea, and the DNA sequence of CM2126 showed high sequence identity to that of P. costantinii (99.5%), and P. grimontii (99.0%). The 16S rDNA sequence of the strain CM2113 showed the highest identity (99.7%) with that of Enterobacter cloacae. The 16S rDNA sequences, the physiological and biochemical analysis suggested that the strain CM2105 belonged to Microbacterium phyllosphaerae, CM2101, CM2121 and CM2126 to Pseudomonas spp., CM2113 to Enterobacter cloacae.

rDNA산물의 정제기술

  • 이승기
    • YAKHAK HOEJI
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    • v.29 no.6
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    • pp.147-155
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    • 1985
  • 이글에서는 예를들어 immunoaffinity chromatography, HPLC및 고압에서 신속하게 분리될수 있게 해주는 새로운 chromatographic media (FPLC)등이 서둘러 개발되고 있는데 본문에섬는 최근에 개발되고 있는 새로운 high resolution 기술을 중심으로 하여 경제성있는 고순도의 rDNA 산물의 회수방법에 대하여 토론하고자 한다. 또한 이들의 정제 방법에 의한 rDNA 산물의 회수에 대한 한계성을 검토하고 최근에 활발히 개발되고 있는 rDNA 조작에 의해 발효산물의 추출및 정제를 용이하게 해줌으로 고수득률과 고순도의 회수를 가능하게 해주는 "upstream processing"기술과 정제 기술의 병용방법에 관하여 집중적으로 토론하고자 한다.

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Pseudomonas 균주에 있어서 R2 Plasmid 획득에 의한 Gamma-ray 내성증강

  • 조봉금
    • Environmental Mutagens and Carcinogens
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    • v.9 no.2
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    • pp.111-121
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    • 1989
  • Ps. aeruginosa 의 DNA repair 기구 결손변이주인 rec-, Hcr- 그리고 R931 plasmid 를 가진 R2 (Carbenicillin, Kanamycin, Streptomycin) plasmid transconjugants 가 R2 Plasmid 획득에 의해서 Gamma선 및 돌연변이제 (4NQO, NTG)에 대해서도 내성을 증강시키는지를 검토함으로써 방사선에 대한 내성화 기구를 해명하고자 했다. 그리고, DNA repair 기구에 작용하는 DNA polymerase I 생산에 관여하는 유전자가 R2 plasmid에 code 되어 있는지를 검토하여 다음과 같은 결과를 얻었다. 1) Ps. aeruginosa PAO균주의 R2 plasmid transconjugants는 R2 plasmid 획득에 의해 자외선, Gamma선 및 돌연변이제에 대한 내성을 부여받았으나 transconjugant 균주에 따라 다른 종류의 내성결과를 얻어졌다.

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High-Throughput DNA Extraction Method for Marker Analysis in Rice Grain (대량의 쌀 시료 분석을 위한 DNA 추출법)

  • Choi, Young-Deok;Lee, Hae-Kwang;Lee, Yun-Suk;Yun, Jeong-Hee;Kim, Su-Jeong;Park, Seong-Whan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.269-273
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    • 2006
  • The study of molecular markers to improve crops largely depends on the availability of rapid and of efficient DNA extraction methods. Here we developed a cheap and convenient method to isolate genomic DNA from rice grains suitable for large-scale microsatellite analysis. We confirmed that the isolated rice DNA is suitable for PCR analysis with STS marker and SNP marker, as well as microsatellite marker. Further, we established high-throughput DNA extraction system in a 96-well plate format which make it possible high-throughput analysis of microsatellite markers with rice grains. This implies that the new method could be a useful tool for other types of marker analysis in large scale.

Phylogenetic Relationship Among Four Species of Korean Oysters Based on Mitochondrial 16S rDNA and COI Gene (미토콘드리아 16S rDNA와 COI유전자에 근거한 한국산 굴류 4종의 유연관계)

  • 이상엽;박두원;안혜숙;김상해
    • Animal Systematics, Evolution and Diversity
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    • v.16 no.2
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    • pp.203-211
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    • 2000
  • Partial mitochondrial 16S rDNA and COI gene were amplified using PCR and sequenced for four species of oysters in Korea. Phylogenetic relationships among them were inferred from their aligned sequences by neighbor-joining method. The sequence comparison data of two mitochondrial genes showed that the genetic distinction between two oyster genera (Crassostreo and Ostrea) was obvious. Phylogenetic analysis based on the nucleotide sequences and A+T percentage of two genes indicates that C. gigas and C. nippona strongly formed a sister group and then C. ariakensis was clustered with the clade although that based on amino acid sequences of COI gene by neighbor-joining method represented different phylogenetic tree.

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