• Title/Summary/Keyword: 16S rRNA analysis

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Bacterial Diversity and Distribution of Cultivable Bacteria Isolated from Dokdo Island (독도 주변의 해수에서 분리한 세균의 다양성과 군집구조 분석)

  • Sung, Hye-Ri;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.263-272
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    • 2010
  • One hundred sixty three strains showing different colony morphological characteristics on different concentration of marine agar (MA) plates were isolated from ambient seawater near Dokdo island. Bacterial diversity and distributions were studied by phylogenetic analysis of the partial 16S rRNA gene sequences. One hundred sixty three strains were partially sequenced and analyzed phylogenetically. They were composed of 5 phyla, of which gamma-proteobacteria (58%), alpha-proteobacteria (20%), bacteriodetes (16%) were predominant. They were affiliated with 90 species. The 16S rRNA sequence similarity of the isolates was in 93.3 to 100 % range to reported sequence data. Thirty six isolates of among them were assumed to be novel species candidates based on similarity analysis of the 16S rRNA gene sequences. Overall, Proteobacteria and Bacteriodetes of the Dokdo coastal sea water showed a high diversity.

Molecular Phylogenetic study of Acila divaricata vigila based on the Partial Sequence of 16S rRNA Gene (민호두조개 (Acila divaricata vigila) 의 16S rRNA 유전자를 기초로 한 분자계통 분류학적 연구)

  • Kim, Bong-Seok;Kang, Se-Won;Jeong, Ji-Eun;Park, Jung-Yeon;Kang, Jung-Ha;Han, Yeon-Soo;Ko, Hyun-Sook;An, Chel-Min;Lee, Jun-Sang;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.27 no.4
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    • pp.395-400
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    • 2011
  • Phylogenetic analyses on the Phylum Mollusks has so far been conducted by many researchers in the world. However, there was no report on taxonomic analysis on Acila divaricata vigila which is belonging to Class Bivalvia, Subclass Protobranchia. In this study, we performed molecular phylogenetic analysis on Acila divaricata vigila using 16S rRNA sequence through maximum likelihood method. As a result, it is clearly divided into the legion of mollusk classification unit (when you zoom in order) and represented to support the current classification in the Phylum Mollusca belong to Class Bivalvia, Subclass Protobranchia, Subclass Pteriomorphia, Subclass Paleoheterodonta, Subclass Heterodonta and Subclass Anomalodesmacea. To our knowledge, this is the first report of molecular phylogenetic analysis on Acila divaricata vigila using 16S rRNA gene and these data suggests that 16S rRNA gene will be useful for analyzing the phylogenetic relationship of Subclass Protobranchia.

A newly developed consensus polymerase chain reaction to detect Mycoplasma species using 16S ribosomal RNA gene

  • Hong, Sunhwa;Park, Sang-Ho;Chung, Yung-Ho;Kim, Okjin
    • Korean Journal of Veterinary Service
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    • v.35 no.4
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    • pp.289-294
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    • 2012
  • Mycoplasmas are highly fastidious bacteria, difficult to culture and slow growing. Infections with Mycoplasma species can cause a variety of problems in living organisms and in vitro cell cultures. In this study, we investigated the usefulness of a genus-specific consensus PCR analysis method to detect Mycoplasma species. The developed consensus primer pairs MycoF and MycoR were designed specifically to amplify the 16S ribosomal RNA gene (rRNA) of Mycoplasma species by the optimized PCR system. The developed consensus PCR system effectively amplified 215 bp of Mycoplasma genus-specific region of 16S rRNA. In conclusion, we recommend this consensus PCR for monitoring Mycoplasma species in animals, human and cell culture system.

Analysis of Microbial Diversity in Makgeolli Fermentation Using PCR-DGGE (PCR-DGGE를 이용한 막걸리발효에서 미생물 다양성 분석)

  • Kwon, Seung-Jik;Ahn, Tae-Young;Sohn, Jae-Hak
    • Journal of Life Science
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    • v.22 no.2
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    • pp.232-238
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    • 2012
  • Kumjungsansung-Makgeolli$^{(R)}$ is a traditional Korean rice wine that is fermented from traditional nuruk and rice. In this study, we performed the PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes to characterize bacterial and fungal diversity during Makgeolli fermentation. The predominant bacteria in the PCR-DGGE profile during Makgeolli fermentation were Lactobacillus spp. (Lactobacillus curvatus, L. kisonensis, L. plantarum, L. sakei, and L. gasseri), Pediococcus spp. (P. acidilactici, P. parvulus, P. agglomerans, and P. pentosaceus), Pantoea spp. (P. agglomerans and P. ananatis), and Citrobacter freundii; these were identified on the base of analysis of 16S rRNA gene sequences. The dominant bacterium during Makgeolli fermentation was L. curvatus. The predominant fungi in PCR-DGGE profile during Makgeolli fermentation were Pichia kudriavzevii, Saccharomyces cerevisiae, Asidia idahoensis, Kluyveromyces marxianus, Saccharomycopsis fibuligera, and Torulaspora delbrueckii, and these were identified on the basis of analysis of 28S rRNA gene sequences. The dominant fungal species during Makgeolli fermentation changed from P. kudriavzevii at 0-2 days incubation to S. cerevisiae at 3-6 days incubation. This study suggests that PCR-DGGE analysis could be a suitable tool for the understanding of microbial diversity and structure during Makgeolli fermentation.

Phylogenetic Analysis of Pectobacterium Species Using the 16S-23S rRNA Intergenic Spacer Regions

  • Kwon, Soon-Wo;Cheun, Meung-Sook;Kim, Sang-Hee;Lim, Chun-Keun
    • The Plant Pathology Journal
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    • v.16 no.2
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    • pp.98-104
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    • 2000
  • For the taxonomic evaluaition, 15 strains of the genus Pectobacterium and Erwinia were analyzed for 16S-23S rDNA intergenic spacer regions (ISRs). These species contained two types of ISRs, large and small ISRs. Large ISRs were on the range of 474-569 bp size, and coding transfer $\textrm{RNA}^{11e}$($\textrm{tRNA}^{11e}$) and $\textrm{tRNA}^{Ala}$. Small ISRs were 354-459 bp in length and coding $\textrm{tRNA}^{Glu}$. The sequence variations of two ISRs among species and strains were very high as compared with 16S rRNA gene sequences. By phylogenetic trees on the basis of two ISRs, Pectobacterium ere differentiated into P. carotovorum-P. cactiaidum group and P. chrysanthemi group. However, the taxonomic position of E. cypripedii and E. rhapontici, which were not clear on taxonomic delineation between Pectobacterium and Erwinia, were not clearly resolved on the basis of ISRs.

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The List of Korean Organisms Registered in the NCBI Nucleotide Database for Environmental DNA Research (환경유전자 연구를 위한 NCBI Nucleotide 데이터베이스에 등록된 국내 생물 목록 현황)

  • Ihn-Sil Kwak;Chang Woo Ji;Won-Seok Kim;Dongsoo Kong
    • Korean Journal of Ecology and Environment
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    • v.55 no.4
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    • pp.352-359
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    • 2022
  • Recently, with the development of genetic technology, interest in environmental DNA (eDNA) to study biodiversity according to molecular biological approaches is increasing. Environmental DNA has many advantages over traditional research methods for biological communities distributed in the environment but highly depends on the established base sequence database. This study conducted a comprehensive analysis of the habitat status and classification at the genus level, which is mainly used in eDNA (12S rRNA, 16S rRNA, 18S rRNA, COI, and CYTB), focusing on Korean registration taxon groups (phytoplankton, zooplankton, macroinvertebrates, and fish). As a result, phytoplankton and zooplankton showed the highest taxa proportion in 18S rRNA, and macroinvertebrates observed the highest ratio in the nucleotide sequence database in COI. In fish, all genes except 18S rRNA showed a high taxon ratio. Based on the Korean registration taxon group, the gene construction of the top 20 genera according to bio density observed that most of the phytoplankton were registered in 18S rRNA, and the most significant number of COI nucleotide sequences were established in macroinvertebrates. In addition, it was confirmed that there is a nucleotide sequence for the top 20 genera in 12S rRNA, 16S rRNA, and CYTB in fish. These results provided comprehensive information on the genes suitable for eDNA research for each taxon group.

Functional Analysis of the Residue 789 in Escherichia coli 16S rRNA and Development of a Method to Select Second-site Revertants (Escherichia coli 16S rRNA의 789 염기의 기능분석 및 이차복귀돌연변이체 발췌를 위한 방법 개발)

  • Kim Jong-Myung;Go Ha-Young;Song Woo-Seok;Ryou Sang-Mi;Lee Kang-Seok
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.156-159
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    • 2006
  • A base substitution was introduced at the position 789 in Escherichia coli 16S rRNA, which was previously identified as an invariant residue for ribosome function and the ability of the mutant ribosomes to translate chloramphenicol acetyltransfernse mRNA was measured by determining the degree of resistance to chloramphenicol of cells expressing these mutant ribosomes. As expected, mutant ribosomes containing a base sub-stitution at the position 789 showed significantly reduced protein-synthesis ability and to identify a functional role played by this residue in protein synthesis, we developed an efficient genetic method to select second-site revertants in 16S rRNA that restore protein-synthesis function to these mutant ribosomes.

Development and Validation of Quick and Accurate Cephalopods Grouping System in Fishery Products by Real-time Quantitative PCR Based on Mitochondrial DNA (두족류의 진위 판별을 위한 Real-time Quantitative PCR 검사법 개발 및 검증)

  • Chung, In Young;Seo, Yong Bae;Yang, Ji Young;Kwon, Ki sung;Kim, Gun Do
    • Journal of Food Hygiene and Safety
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    • v.33 no.4
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    • pp.280-288
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    • 2018
  • In this study, an approach for the analysis of the five cephalopod species (octopus, long-arm octopus, squid, wet-foot octopus, beka squid) consumed in the Republic of Korea is developed. The samples were collected from the Southeast Asian countries Thailand, Indonesia, Vietnam, and China. The SYBR-green-based real-time qPCR method, based on the mitochondrial DNA genome of the five cephalopods was developed and validated. The intergroup variations in the mitochondrial DNA are evident in the bioinformatic analysis of the mitochondrial genomic DNA sequences of the five groups. Some of the highly-conserved and slightly-variated regions are identified in the mitochondrial cytochrome-c-oxidase subunit I (COI) gene, 16s ribosomal RNA (16s rRNA) gene, and 12s ribosomal RNA (12s rRNA) gene of these groups. To specify each five cephalopod groups, specific primer sets were designed from the COI, 16s rRNA and 12s rRNA regions. The specific primer sets amplified the DNA using the SYBR-green-based real-time PCR system and 11 commercially secured animal tissues: Octopus vulgaris, Octopus minor, Todarodes pacificus, Dosidicus gigas, Sepia esculenta, Amphioctopus fangsiao, Amphioctopus aegina, Amphioctopus marginatus, Loliolus beka, Loligo edulis, and Loligo chinensis. The results confirmed by a conveient way to calculate relative amplification levels between different samples in that it directly uses the threshold cycles (Ct)-value range generated by the qPCR system from these samples. This genomic DNA-based molecular technique provides a quick, accurate, and reliable method for the taxonomic classification of the animal tissues using the real-time qPCR.

Molecular diversity of endobacterial communities in edible part of King oyster mushroom (Pleurotus eryngii) based on 16S rRNA (16S rRNA 기초 새송이 버섯(Pleurotus eryngii)의 식용가능 부위 내생세균 군집 다양성)

  • Lee, Choung Kyu;Haque, Md. Azizul;Choi, Byoung Rock;Lee, Hee Yul;Hwang, Chung Eun;Ahn, Min Ju;Cho, Kye Man
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.148-155
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    • 2015
  • The diversity of endobacteria in the edible part (cap and stipe) king oyster mushroom (Pleurotus eryngii) was investigated using 16S rRNA sequence analysis. The bacterial 16S rRNA libraries were constructed from the body cap (BC) and the body stipe (BS) of the king oyster mushroom. The twenty sequenced BC clones were divided into four groups and the largest group was affiliated with the Firmicutes (40% of clones). While, the twenty sequenced BS clones could be divided into six groups and the largest group was affiliated with the Actinobacteria (40% of clones). The predominant bacterial family from both the cap and stipe of the mushroom was corresponded with the Gram positive bacteria (62.5%).

Phylogenetic Relationships among Allium subg. Rhizirideum Species Based on the Molecular Variation of 5S rRNA Genes

  • Do, Geum-Sook;Seo, Bong-Bo
    • Animal cells and systems
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    • v.4 no.1
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    • pp.77-85
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    • 2000
  • This study has demonstrated the molecular variation of 5S rRNA genes in 15 Allium subgenus Rhizirideum and 1 Allium subg. Allium. For cloning of the 5S rRNA genes, PCR products were obtained from amplification with oligonucleotide primers which were derived from the conserved coding region of 5S rRNA genes. These amplified PCR products were cloned and identified by FISH and sequence analysis. The 5S rRNA loci were primarily located on chromosomes 5 and/or 7 in diploid species and various chromosomes in alloploid species. The size of the coding region of 5S rRNA genes was 120 bp in all the species and the sequences were highly conserved within Allium species. The sizes of nontranscribed spacer (NTS) region were varied from 194 bp (A. dektiude-fustykisum, 2n=16) to 483 bp (A. sativum). Two kinds of NTS regions were observed in A. victorialis var. platyphyllum a diploid, A. wakegi an amphihaploid, A. sacculiferum, A. grayi, A. deltoide-fistulosum and A. wenescens all allotetraploids, while most diploid species showed only one NTS region. The species containing two components of NTS region were grouped with different diploid species in a phylogenetic tree analysis using the sequences of 5S rRNA genes and adjacent non-coding regions.

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