• Title/Summary/Keyword: 16S-rRNA

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Use of 16S-23S rRNA Intergenic Spacer Region for Rapid Detection of Vibrio fluvialis (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio fluvialis의 검출)

  • 강현실;허문수;이제희
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.77-85
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    • 2003
  • We have examined the 16S-23S rRNA intergenic spacer regions (ISR) of Vibrio fluvialis. ISRs were PCR amplified, cloned into a plasmid vector and then sequenced. As results of ISR nucleotide sequence analysis, total of 6 clones were isolated depending on the size. The clones were different in both the number and the composition of the tRNA genes, and were designated ISR-A, ISR-E, ISR-El, ISR-lA, ISR-EKV, ISR-EKAV. ISR-A contains $tRNA^{Ala}$; ISR-lA, $tRNA^{Ile}$-$tRNA^{Ala}$; ISR-EKV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Val}$;ISE-EKAV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Ala}$-$tRNA^{Val}$; ISR -E and E1, $tRNA^{GIu}$ clusters. ISR-EKV was shown to be a minor type out of the six ISR types and showed a very limited homology between ISR-EKV from V, fluvialis and ISRa from other Vibrio species. Therefore ISR-EKV sequence was used to design species-specific primers to detect V, fiuvialis from other Vibrio species by PCR reaction. The specificity of the primers was examined using genomic DNA of other Vibrios as templates for PCR reaction. The result showed that PCR can be a useful method to detect V. fluvialis among Vibrio species in a single PCR reaction.

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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Archaeal Communities in Mangrove Soil Characterized by 16S rRNA Gene Clones

  • Yan, Bing;Hong, Kui;Yu, Zi-Niu
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.566-571
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    • 2006
  • An archaeal 16S rRNA gene library was constructed from mangrove soil. Phylogenetic analysis revealed archaea in mangrove soil including the Crenarchaeota (80.4%) and Euryarchaeota (19.6%) phyla. The archaeal community in mangrove soil appears to be a mixture of organisms found in a variety of environments with the majority being of marine origin.

Phytoplasma specific primer for detection of jujube witches′ broom group(16SrV) in Korea and China

  • Sangsub Han;Lee, Sanghun;Mengjun Liu;Byeongjin Cha
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.136.2-137
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    • 2003
  • In order to diagnose and differentiate jujube witches' broom (JWB) phytoplasma rapidly, oligonucleotide primer pair, 16Sr(V) F/R, for polymerase chain reactions (PCRs) was designed on the basis of 165 rRNA sequences of JWB phytoplasma. The PCR employing phytoplasma universal primer pair P1/P7 consistently amplified DNA in all tested phytoplasma isolates. But no phytoplasma DNA was detected in healthy jujube seedlings. The nested PCR, the primer pair 16S(V) F/R, about 460 bp fragment, amplified DNA in all tested JWB and related phytoplasmas including LiWB phytoplasma of the 165 rRNA group V, but no DNA amplification was detected from other phytoplasma strains such as group 16SrI (Aster yellows) and group 16SrⅩII (Stolbur group) phytoplasmas in which mulberry dwarf phytoplasma and chrysanthemum witches broom phytoplasma are belonged to, respectively The same results were obtained from both Korean- and Chinese-isolates of JWB. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16S rRNA group V specific primer pair 16S(V) F/R could detect group V phytoplasma rapidly and easily, in particular JWB phytoplasma.

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Down-regulation of the cyclin E1 oncogene expression by microRNA-16-1 induces cell cycle arrest in human cancer cells

  • Wang, Fu;Fu, Xiang-Dong;Zhou, Yu;Zhang, Yi
    • BMB Reports
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    • v.42 no.11
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    • pp.725-730
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    • 2009
  • Cyclin E1 (CCNE1), a positive regulator of the cell cycle, controls the transition of cells from G1 to S phase. In numerous human tumors, however, CCNE1 expression is frequently dysregulated, while the mechanism leading to its dysregulation remains incompletely defined. Herein, we showed that CCNE1 expression was subject to post-transcriptional regulation by a microRNA miR-16-1. This was evident at protein level of CCNE1 as well as its mRNA level. Further evident by dual luciferase reporter assay revealed that two evolutionary conserved binding sites on 3' UTR of CCNE1 were the direct functional target sites. Moreover, we showed that miR-16-1 induced G0/G1 cell cycle arrest by targeting CCNE1 and siRNA against CCNE1 partially phenocopied miR-16-1-induced cell cycle phenotype whereas substantially rescued anti-miR-16-1- induced phenotype. Together, all these results demonstrate that miR-16-1 plays a vital role in modulating cellular process in human cancers and indicate the therapeutic potential of miR-16-1 in cancer therapy.

수계 생태계에서의 세균 군집 구조의 분자생물학적 분석

  • 이동훈;김상종
    • Korean Journal of Microbiology
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    • v.33 no.1
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    • pp.55-65
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    • 1997
  • 16S rRNA를 분석한 연구들은 자연 생태계에서 추출한 핵산을 이용하여 16rRNA 유전자의 염기서열을 분석하거나 특정 DNA probe를 이용한 hybridization 실험이 주류를 이루어 왔다. 특히 PCR 기법이 개발됨에 따라 적은 양의 시료를 대량으로 손쉽게 증폭시킬 수 있어 다양한 분야에 응용되고 있다. 세균 군집의 구조를 이해하는데 있어서 PCR 방법의 적용 대상은 주로 16S rRNA 유전자의 염기서열 해독분야이며 해양 생태계를 대상으로 많은 연구 결과가 보고되었다(11,13,21,26). 한편 자연 생태계의 개별적 미생물 분류룬들을 검출하기 위한 특정 oligonucleotide probe의 개발방법들은 미생물 군집의 유전적 다양성에 대한 정보 파악 이외에 배양이 어려운 혐기성 세균과 같은 특정 세균들의 동정에도 이용되고 있다(3,24,55). 본고에서는 세균 군집의 구조와 다양성을 연구하는데 적용 가능한 rRNA 분석방법들을 수계 생태계를 중심으로 살펴보고자 한다.

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16S rRNA gene-based sequencing of cucumber (Cucumis sativus L.) microbiota cultivated in South Korea (16S rRNA 유전자 염기서열 분석에 기반한 국내 재배 오이의 상재균총 분석)

  • Seo, Dong Woo;Kim, Seung Min;Lee, Heoun Reoul;Yum, Su-jin;Jeong, Hee Gon
    • Korean Journal of Food Science and Technology
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    • v.53 no.3
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    • pp.334-343
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    • 2021
  • Various vegetables, including cucumbers, have a high probability of foodborne illness because they are usually eaten raw. In this study, we analyzed the 16S rRNA gene sequences of the cucumber (Cucumis sativus L.) microbiota. The diversity indices of cucumber cultivated in May were higher than in cucumber cultivated in November. At the phylum level, Proteobacteria, Firmicutes, and Actinobacteria were predominant. The classes generally comprised Gammaproteobacteria, Bacilli, Alphaproteobacteria, and Actinobacteria. At the genus level, the proportions of Aureimonas, Escherichia, and Microbacterium in samples from May were relatively high, whereas Enterococcus, Pseudomonas, and Rhizobium accounted for a higher proportion in samples from November. Moreover, it is noteworthy that potential pathogenic genera such as Acinetobacter, Aerococcus, Aureimonas, Enterobacter, Enterococcus, Escherichia, Pantoea, Pseudomonas, and Staphylococcus were detected. Although further studies on the characteristics of potential pathogens are required, our results can be used to improve the food safety of vegetables.

Occurrence of Petunia Flattened Stem Caused by Phytoplasma

  • Chung, Bong-Nam;Huh, Kun-Yang
    • The Plant Pathology Journal
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    • v.24 no.3
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    • pp.279-282
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    • 2008
  • This study describes a phytoplasmal disease occurring in Petunia leaves grown in the glasshouse of the National Horticultural Research Institute, Suwon, Korea. Abnormal growth like flattened stem with flower malformation or phyllody was observed from the plant. The DNA extracted from the diseased leaves was amplified using a universal primer pair of P1/P6 derived from the conserved 16S rRNA gene of Mollicutes giving the expected polymerase chain reaction(PCR) product of 1.5 kb. In the nested PCR assays, the expected DNA fragment of 1.1 kb was amplified with the specific primer pair R16F1/R16R1 that was designed on the basis of aster yellows(AY) phytoplasma 16S rDNA sequences. The 1.1 kb PCR products were cloned and nucleotide sequences were determined, and the sequences of the cloned 168 rRNA gene were deposited in the GenBank database under the accession no. of EU267779. Analysis of the homology percent of the 168 rDNA of PFS-K showed the closest relationship with Hydrangea phyllody phytoplasma(AY265215), Brassica napus phytoplasma(EU123466) and AY phytoplasma CHRY(AY180956). Phytoplasma isolated from the diseased Petunia was designated as Petunia flat stem phytoplasma Korean isolate(PFS-K) in this study. Flattened stem occurring in Petunia was confirmed as infection of AY group of phytoplasma by determination of 16S rRNA gene sequences of phytoplasma and microscopic observation of phytoplasma bodies. This is the first report on the phytoplasmal disease in Petunia in Korea.

Bacterial Diversity in a Korean Traditional Soybean Fermented Foods (Doenjang and Ganjang) by 16S rRNA Gene Sequence Analysis

  • Cho, Kye-Man;Seo, Weon-Taek
    • Food Science and Biotechnology
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    • v.16 no.2
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    • pp.320-324
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    • 2007
  • The bacterial diversity in Korean soybean-fermented foods was investigated using a PCR-based approach. 16S rRNA sequences were amplified and cloned from two different soybean-fermented foods such as doenjang (soybean paste), and ganjang (soybean sauce). Staphylococcus equorum (60.6%), Tetragenococcus halophila (21.2%), Leuconostoc mesenteroides (9.1%), Lactobacillus sakei (6.1%), and Bacillus subtilis (3.0%) were detected among clones isolated from soybean paste samples and Halanaerobium sp. (37.5%), Halanaerobium fermentans (37.5%), T. halophila (12.5%), Staphylococcus sp. (6.3%), S. equorum (3.1%), and B. subtilis (3.1%) were detected among clones isolated from soybean sauce. Our approach revealed different bacterial distributions and diversity from those previously obtained using culture-dependent methods.

Prevalence and molecular characteristics of 16s rRNA methylase gene rmtB in amikacin resistant Escherichia coli isolated from South Korea

  • Belaynehe, Kuastros Mekonnen;Won, Ho Geun;Yoon, In Joong;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.59 no.3
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    • pp.157-160
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    • 2019
  • The production of rmtB-encoded 16S rRNA methylases has emerged as a novel mechanism promoting high-level resistance toward aminoglycosides in Gram-negative bacteria. Between 2015 and 2017, 636 distinct commensal Escherichia (E.) coli isolates were collected from different farms in South Korea to determine the prevalence and molecular characteristics of rmtB. The positive rates of rmtB between all the isolates and amikacin-resistant isolates were 1.1 and 100%, respectively. High-level aminoglycoside resistance could be transferred by conjugation from rmtB-positive donors to higher amikacin-resistance efficacies. This is the first report of 16S rRNA methylase-encoding genes in E. coli isolated from food-producing animals in Korea.