• Title/Summary/Keyword: 16S ribosomal RNA

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Monitoring of Commercial Products Sold on Sushi Buffet Restaurants in South Korea using DNA Barcode Information (국내 대형 초밥 뷔페에서 사용되는 수산물의 원재료 모니터링 연구)

  • Kang, Tae Sun
    • Journal of Food Hygiene and Safety
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    • v.35 no.1
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    • pp.45-50
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    • 2020
  • In this study, seafood products (n=26) sold on sushi buffet restaurants in the city of Wonju were monitored by analyzing sequences of DNA barcode markers (cytochrome c oxidase subunit I and 16S ribosomal RNA genes). NCBI BLAST database was screened with the barcode sequences analyzed as a query for species identification. The BLAST search revealed that fifteen samples (58%) analyzed were consistent with their labeling information; however, the ingredients used in seven samples (27%) were not compliant with their label information. In the case of these mislabeled products, ingredients for sutchi catfish sushi and cherry bass sashimi were identified as Pangasianodon hypophthalmus and Lampris guttatus, respectively. For Japanese flying-fish roe sushi and Pacific herring roe sushi, roe of Mallotus villosus was used as an ingredient. Amphioctopus fangsiao and A. membranaceus were used in octopus sushi and soybean-marinated squid products, respectively. This monitoring result can contribute to the protection of consumer rights and the reduction of fraudulent practices in the food industry.

Phylogeneitc Analysis of Fluorescent Pseudomonas spp. Isolated from the Cultivated Mushrooms on the Basis of ITS I Region (버섯에서 분리한 형광성 Pseudomonas spp. 의 ITS I 영역 분석에 의한 계통 분류)

  • 고승주;고승주;강희완;전명숙;류진창
    • Korean Journal Plant Pathology
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    • v.14 no.4
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    • pp.350-357
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    • 1998
  • A total of 12 strains of fluorescent Pseudomonas isolated from the cultivated mushrooms such as Agaricus bisporus and Pleurotus ostreatus were collected. They consisted of pathogenic Pseudomonas spp. and epiphytic Pseudomonas spp. of the cultivated mushroom. To analyze the phylogenetic relationship of these strains, ITS I region, the 16S-23S intergenic spacer region in the ribosomal RNA (rRNA) operon, was cloned and sequenced. The spacer regions of these strains were 495∼527 nucleotides in length and contained the genes encoding isoleucine-tRNA (tRNAIle) and alanine-tRNA (tRNAAla). The reciprocal homologies of each ITS I sequence among these strains were in the range of 84.2%∼98.8%. According to the analysis of ITS I sequences, the fluorescent Pseudomonas spp. were phylogenetically classified into three clusters. Cluster I consisted of Pseudomonas fluorescens, P. tolaasii, P. gingeri’, and P.‘reactans’(WLRO). Cluster II comprised Pseudomonas fluorescens biovar C and F. Cluster III composed P. agarici. Cluster I and II could be classified into P. fluorescens complex. P. agarici formed an independent taxon clearly separable from P. florescens complex.

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Phylogenetic analysis of Locusta migratoria (Orthoptera: Acridae) in Haenam-gun, Jeollanam-do, Korea using Two Mitochondrial Genes (마이토콘드리아 유전자 2개를 이용한 대한민국 전라남도 해남군 발생 풀무치 Locusta migratoria (메뚜기목: 메뚜기과)의 계통분석)

  • Kim, Young-Ha;Jung, Jin-Kyo;Lee, Gwan-Seok;Koh, Young-Ho
    • Korean journal of applied entomology
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    • v.55 no.4
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    • pp.459-464
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    • 2016
  • An outbreak of the migratory locust, Locusta migratoria, in the environment-friendly reclaimed plantations of forage crops in Sanyimyeon, Haenam-gun, Jellanam-do, Korea in August 2014 caused severe damages to various crops. Owing to its first occurrence in the Korean history, the causes underlying the outbreak and phase-transition of the migratory locust were not known. It is critical to establish the genetic relationship of the migratory locust in Sanyimyeon, Haenam-gun with the other previously reported strains in the world in order to understand the mechanisms responsible for its outbreak. The gene sequences of the 16S ribosomal RNA (rRNA) and displacement-loop (D-loop) of the mitochondria of various regional species of the migratory locust were used to perform the phylogenetic analysis. Our results suggested that the migratory locusts in Sanyimyeon, Haenam-gun are closely related with the Eurasian strains of the northern lineage. In future, these two mitochondrial genes can be used for elucidating the genetic population structures in migratory locusts in various regions. In addition, the sequence information of these genes can be used to enhance our understanding of the genetic basis of the outbreak of migratory locusts.

Antagonism against Helicobacter Pylori and Proteolysis of Lactobacillus Helveticus CU631 and Strain Identification

  • Yoon, Y.H.;Won, B.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1057-1065
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    • 2002
  • The antagonistic activities of 30 strains of lactobacilli against Helicobacter pylori were determined and Lactobacillus helveticus CU631 has been selected as the strain which possesses the strongest inhibitory effect in the disc diffusion assay showing inhibition zone diameter of $10{\pm}1.5mm$, whereas those of L. plantarum and L. fermentum have been shown to be $4.0{\pm}0.6mm$. H. pylori G88016 revealed the highest vacuolating toxin producing activity among the 8 strains, the inhibitory activity of L. helveticus CU631 in vacuolating toxin producing activity of H. pylori manifested in the co-culture of two strains and in the 5:5 mixture of supernatant of the two strains. Both L. helveticus CU631 and cell free culture supernatant had a strong inhibitory activities in urease and cytotoxin producing activities of H. pylori NCTC11637 and CJH12. An accelerated proteolytic activity of water soluble peptides by L. helveticus CU631 during the refrigeration storage has been manifested in the cream cheese. DNA seqences of 16S-23S ribosomal RNA spacer region showed typical pattern among the various strains of L. helveticus, which could be used in the identification of L. helveticus CU 631.

Intrageneric Relationships of Trichoderma Based on Internal Transcribed Spacers and 5.8S rDNA Nucleotide Sequences

  • Kim, Gi-Young;Lee, Goang-Jae;Ha, Myung-Gyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.28 no.1
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    • pp.11-16
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    • 2000
  • The nucleotide sequences of the internal transcribed spacer (ITS) regions of the ribosomal DNA including the 5.8S ribosomal RNA gene (rDNA) have been determined for 11 species in order to analyze their intrageneric relationships. The total length of these sequences ranged from 530 nucleotides for Trichoderma reesei KCTC 1286 to 553 nucleotide for Trichoderma koningii IAM 12534. Generally speaking, the length of ITS1 region was about 30 nucleotides longer than that of the ITS2 region. Also, the sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites were also found. Thus, a neighbor-joining tree was constructed using the full sequence data of the ITS regions and the 5.8S rDNA. The Trichoderma genus used to be grouped on the basis of the morphological features and especially the shape of phialides needs to be reexamined. The phylogenetic tree displayed the presence of monophylogeny in the species of Trichoderma. Therefore, it was difficult to distinguish the intrageneric relationships in the Trichoderma genus.

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Studies on the mycoplasmal pneumonia in slaughter pigs. 2. Isolation of mycoplasmas from lung tissues and identification of isolates by nested-PCR technique (도축돈의 마이코플라즈마성 폐렴에 관한 연구 2. 폐조직에서의 균분리와 nested-PCR방법에 의한 동정)

  • Lim, Young-Taek;Seok, Ho-Bong
    • Korean Journal of Veterinary Research
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    • v.42 no.2
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    • pp.225-229
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    • 2002
  • We report that mycoplasma organisms from lung tissues of slaughter pigs were identified to genes fragments with references use of nested-PCR technique(nPCR). Seven strains of mycoplasma species were isolated from 70 lung tissues. The organisms were detected by in vitro amplification of 16S rRNA and 23S rRNA genes. Nucleotide sequences of the spacer between 16S and 23S in the ribosomal RNA operons of mycoplasma were identified by the analysis of products from the nested PCR. Four common PCR primers, MhF1, MhF2 MhR1 and MhR2, were designed by analysis between these sequences by first amplified with F1, R1 and second with F2, R2, respectively. Specific amplification of the spacer region for reference strains of M. hyopneumoniae, M. hyorhinis, M. flocculare were confirmed by first round of PCR in which the traduced fragments of 690bp, 460bp, 630bp. But amplications of second round was changed to 240bp, 210bp, 230bp, respectively. Three different strains (M. hyopneumoniae:4, M. hyorhinis:2, M. flocculare:1) were detected by the nested-PCR technique. The results suggest that the detection of swine mycoplasma by n-PCR can be analyzed the nucleotide sequences between rRNA operons and homology study.

Diversity Census of Fecal Microbiome in Horses (말 분변 내 마이크로바이옴 다양성 조사)

  • Lee, Seul;Kim, Minseok
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.157-165
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    • 2019
  • This study was conducted to analyze the diversity census of fecal microbiome in horses using meta-analysis of equine 16S rRNA gene sequences that are available in the Ribosomal Database Project (RDP; Release 11, Update 5). The search terms used were "horse feces (or faeces)" and "equine feces (or faeces)". A total of 842 sequences of equine feces origin were retrieved from the RDP database, where 744 sequences were assigned to 10 phyla placed within Domain Bacteria. Firmicutes (n = 391) and Bacteroidetes (n = 203) were the first and the second dominant phyla, respectively, followed by Verrucomicrobia (n = 58), Proteobacteria (n = 30) and Fibrobacteres (n = 24). Clostridia (n = 319) was the first dominant class placed within Bacteroidetes while Bacteroidia (n = 174) was the second dominant class placed within Bacteroidetes. The remaining 98 sequences were assigned to phylum Euryarchaeota placed within Domain Archaea, where 74 sequences were assigned to class Methanomicrobia. The current results will improve understanding of the diversity of fecal microbiome in horses and may be used to further analyze equine fecal microbiome in future studies.

Simultaneous Quantification of Cyanobacteria and Microcystis spp. Using Real-Time PCR

  • Oh, Kyoung-Hee;Jeong, Dong-Hwan;Shin, Seung-Hee;Cho, Young-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.248-255
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    • 2012
  • In order to develop a protocol to quantify cyanobacteria and Microcystis simultaneously, the primers and probe were designed from the conserved regions of 16S rRNA gene sequences of cyanobacteria and Microcystis, respectively. Probe match analysis of the Ribosomal Database Project showed that the primers matched with over 97% of cyanobacterial 16S rRNA genes, indicating these can be used to amplify cyanobacteria specifically. The TaqMan probe, which is located between two primers, matched with 98.2% of sequences in genus GpXI, in which most Microcystis strains are included. The numbers of cyanobacterial genes were estimated with the emission of SYBR Green from the amplicons with two primers, whereas those of Microcystis spp. were measured from the fluorescence of CAL Fluor Gold 540 emitted by exonuclease activity of Taq DNA polymerase in amplification. It is expected that this method enhances the accuracy and reduces the time to count cyanobacteria and potential toxigenic Microcystis spp. in aquatic environmental samples.

Trimming conditions for DADA2 analysis in QIIME2 platform

  • Lee, Seo-Young;Yu, Yeuni;Chung, Jin;Na, Hee Sam
    • International Journal of Oral Biology
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    • v.46 no.3
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    • pp.146-153
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    • 2021
  • Accurate identification of microbes facilitates the prediction, prevention, and treatment of human diseases. To increase the accuracy of microbiome data analysis, a long region of the 16S rRNA is commonly sequenced via paired-end sequencing. In paired-end sequencing, a sufficient length of overlapping region is required for effective joining of the reads, and high-quality sequencing reads are needed at the overlapping region. Trimming sequences at the reads distal to a point where sequencing quality drops below a specific threshold enhance the joining process. In this study, we examined the effect of trimming conditions on the number of reads that remained after quality control and chimera removal in the Illumina paired-end reads of the V3-V4 hypervariable region. We also examined the alpha diversity and taxa assigned by each trimming condition. Optimum quality trimming increased the number of good reads and assigned more number of operational taxonomy units. The pre-analysis trimming step has a great influence on further microbiome analysis, and optimized trimming conditions should be applied for Divisive Amplicon Denoising Algorithm 2 analysis in QIIME2 platform.

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.