• Title/Summary/Keyword: 16S rRNA analyzing method

Search Result 7, Processing Time 0.022 seconds

The detection of subgingival plaque microflora using 16S rRNA analysis in Korean adult periodontitis (한국인 성인성 치주염 환자에서 16S rRNA 분석을 이용한 치은연하치태 세균 분포도 조사)

  • Park, Seong-Hee;Kim, So-Young;Choi, Seong-Ho;Chai, Jung-Kiu;Kim, Chong-Kwan;Cho, Kyoo-Sung
    • Journal of Periodontal and Implant Science
    • /
    • v.28 no.4
    • /
    • pp.691-703
    • /
    • 1998
  • The 16S rRNA analyzing method is a bacterial identification method that is useful in identifying bacteria which is difficult to do by other means. The following 7 types of bacteria which are Treponema, A. actinomycetemcomitans, P. gingivalis, Fusobacterium, B. forsythus, P. intermedia, P. micros were evaluated in order to study their distribution among patients with adult periodontitis. The 16S rRNA analyzing method was used to compare bacterial distribution among 3 groups. Subgingival plaque acquired from the affected sites(pocket depth ${\geq}6mm$) of 29 patients with adult periodontitis were grouped as the experimental group while plaque from the non-affected sites(pocket depth ${\leq}3mm$) were grouped as control 2 and finally plaque acquired from students with healthy periodontal tissues were grouped as control 1. The results are as follows ; 1. The distribution of Treponema was 12.5% for control 1, 21.4% for control 2 and 75.4% for the experimental group. For A. actinomycetemcomitans the distribution was 0.5%, 19.0%, 44.4% in respect to the order of groups mentioned above. P.gingivalis showed 10.5%, 43.1%, 94.0% distribution, Fusobacterium 33.0%, 48.3%, 81.0% distribution, B. forsythus 9.5%, 17.2%, 65.9% distribution, P. intermedia 1.0%, 12.1%, 26.3% distribution and finally P. micros 5.0%, 19.0%, 48.7% respectively. In all 7 types of bacteria, the experimental group showed higher bacterial distribution compared to the other two groups with statistically significant difference. 2. In the case of Treponema, A. actinomycetemcomitans, gingivalis,Fusobacterium, B. forsythus, P. intermedia, P. micros showed significant difference between control 1 and 2. These results suggest that the 16S rRNA analyzing method which was applied on Koreans for the first time could be utilized and useful in finding potential pathogens of periodontal disease.

  • PDF

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
    • /
    • v.27 no.4
    • /
    • pp.395-400
    • /
    • 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.

Rapid Preparation of Total Nucleic Acids from E. coli for Multi-purpose Applications

  • Cheng, Lin;Li, Tai-Yuan;Zhang, Yi
    • BMB Reports
    • /
    • v.37 no.3
    • /
    • pp.351-355
    • /
    • 2004
  • Separate protocols are commonly used to prepare plasmid DNA, chromosomal DNA, or total RNA from E. coli cells. Various methods for the rapid preparation of plasmid DNA have been developed previously, but the preparation of the chromosomal DNA and total RNA are usually laborious. We report here a simple, fast, reliable, and cost-effective method to extract total nucleic acids from E. coli by direct lysis of the cells with phenol. Five distinct and sharp bands, which correspond to chromosomal DNA, plasmid DNA, 23S rRNA, 16S rRNA, and a mixture of small RNA, were observed when analyzing the prepared total nucleic acids on a regular 1-2% agarose gel. The simple and high-quality preparation of the total nucleic acids in a singe tube allowed us to rapidly screen the recombinant plasmid, as well as to simultaneously monitor the change of the plasmid copy number and rRNA levels during the growth of E. coli in the liquid medium.

Study on the Genetic Variation of the Mitochondrial DNA in the Beet Armyworm, Spodoptera exigua (H bner), Using PCR-RFLP (PCR-RFLP를 이용한 파방나방 (Spodoptera exigua(H bner)) 미토콘트리아 DNA의 유전변이 연구)

  • 김용균;이명렬;정충렬
    • Korean journal of applied entomology
    • /
    • v.37 no.1
    • /
    • pp.23-30
    • /
    • 1998
  • Restriction fragment length polymorphism (RFLP) of a DNA has been a useful tool for analyzing genetic variation. This research was performed to establish an RFLP analytic method on the mitochondrial DNA (mtDNA) of the beet armyworm, Spodoptera exigua (Hiibner). To do this, total size of the mtDNA was measured and polymerase chain reaction (PCR) primers were selected. Its mitochondrial genome size was ca. 16kb. From a serial PCR test of 29 primers refered to the compilation of Simon et al. (1994), 22 primers were selected to amplify its mtDNA fragments. These primers resulted in short (300-700 bp) or long (1000-2000 bp) DNA products which represented a total or partial sequence of each of CO-I, CO-11, Cyt-B, ND-1, 12s rRNA, 16s rRNA, and some tRNAs. PCR-RFLP was performed in some variable mtDNA regions with 8 kinds of 4bp recognizing restriction enzymes. Different populations from Andong, Kyungsan, and Sunchun did not show any restriction site polymorphisms but had some length variation in certain regions of mtDNA.

  • PDF

Analysis of Stomach Contents of Marine Orgnaisms in Gwangyang Bay and Yeosu Fish Market Using DNA Metabarcoding (DNA 메타바코딩을 이용한 광양만 및 어시장 해양 생물 위 내용물 분석)

  • Gun Hee Oh;Yong Jun Kim;Won-Seok Kim;Cheol Hong;Chang Woo Ji;Ihn-Sil Kwak
    • Korean Journal of Ecology and Environment
    • /
    • v.55 no.4
    • /
    • pp.368-375
    • /
    • 2022
  • Gut contents analysis is essential to predict the impact of organisms on food source changes due to variations of the habitat environment. Previous studies of gut content analysis have been conducted using traditional methods, such as visual observation. However, these studies are limited in analyzing food sources because of the digestive process in gut organ. DNA metabarcoding analysis is a useful method to analyze food sources by supplementing these limitations. We sampled marine fish of Pennahia argentata, Larimichthys polyactis, Crangon affinis, Loligo beka and Sepia officinalis from Gwangyang Bay and Yeosu fisheries market for analyzing gut contents by applying DNA metabarcoding analysis. 18S rRNA v9 primer was used for analyzing food source by DNA metabarcoding. Network and two-way clustering analyses characterized the relationship between organisms and food sources. As a result of comparing metabarcoding of gut contents for P. argentata between sampled from Gwangyang Bay and the fisheries market, fish and Copepoda were analyzed as common food sources. In addition, Decapoda and Copepoda were analyzed as common food sources for L. polyactis and C. affinis, respectively. Copepoda was analyzed as the primary food source for L. beka and S. officinalis. These study results demonstrated that gut contents analysis using DNA metabarcoding reflects diverse and detailed information of biological food sources in the aquatic environment. In addition, it will be possible to provide biological information in the gut to identify key food sources by applying it to the research on the food web in the ecosystem.

Dual Application of p-Nitrophenol Alkanoate-Based Assay for Soil Selection and Screening of Microbial Strains for Bioplastic Degradation

  • Nara Shin;Jinok Oh;Suwon Kim;Yeda Lee;Yuni Shin;Suhye Choi;Shashi Kant Bhatia;Yung-Hun Yang
    • Journal of Microbiology and Biotechnology
    • /
    • v.34 no.7
    • /
    • pp.1530-1543
    • /
    • 2024
  • With an increase in the commercialization of bioplastics, the importance of screening for plastic-degrading strains and microbes has emerged. Conventional methods for screening such strains are time-consuming and labor-intensive. Therefore, we suggest a method for quickly and effectively screening plastic-degrading microbial strains through dual esterase assays for soil and isolated strains, using p-nitrophenyl alkanoates as substrates. To select microbe-abundant soil, the total amount of phospholipid fatty acids (PLFAs) included in each soil sample was analyzed, and esterase assays were performed for each soil sample to compare the esterase activity of each soil. In addition, by analyzing the correlation coefficients and sensitivity between the amount of PLFAs and the degree of esterase activity according to the substrate, it was confirmed that substrate pNP-C2 is the most useful index for soil containing several microbes having esterase activity. In addition, esterase assays of the isolated strains allowed us to select the most active strain as the degrading strain, and 16S rRNA results confirmed that it was Bacillus sp. N04 showed the highest degradation activity for polybutylene succinate (PBS) as measured in liquid culture for 7 days, with a degradation yield of 99%. Furthermore, Bacillus sp. N04 showed degradation activity against various bioplastics. We propose the dual application of p-nitrophenyl alkanoates as an efficient method to first select the appropriate soil and then to screen for plastic-degrading strains in it, and conclude that pNP-C2 in particular, is a useful indicator.

Positive effects of grazing on blood components and intestinal microbiota in growing horses

  • Ji Hyun Yoo;Jong An Lee;Jae Young Choi;Sang Min Shin;Moon Cheol Shin;Hyeon Ah Kim;Yong Jun Kang;Hee Chung Ji;In Cheol Cho;Byoung Chul Yang
    • Korean Journal of Agricultural Science
    • /
    • v.49 no.4
    • /
    • pp.963-971
    • /
    • 2022
  • Production of high-quality horses is important to make the horse industry grow. Grazing during the growing period can be an important factor affecting the production of high-quality horses. The objective of this study was to determine the effects of grazing on growing horses by analyzing their blood components and intestinal microbiota. Twelve growing horses for evaluating blood components and ten growing horses for evaluating intestinal microbiota were raised for about seven months and separated by two treatments: grazing vs. stable. Complete blood count, blood chemistry, and creatine kinase levels were analyzed as blood components and a 16s rRNA gene sequence analysis was performed to analyze intestinal microbiota. Calcium ions tended to be lower in the group with grazing treatment. Alkaline phosphatase and creatine kinase tended to be higher in the group with grazing treatment. These results indicate that grazing can provide horses with more exercise than staying in stables. At the phylum level, Firmicutes/Bacteroidetes ratios in grazing and stable groups were 4.2 and 6.5, respectively. Because various studies have reported that a. high Firmicutes/Bacteroidetes ratio indicates obesity, the method of raising horses might affect their physical ability. At the species level, rates of Clostridium butyricum in grazing and stable groups were 3.2% and 13.1%, respectively. Some strains of C. butyricum can cause several diseases such as botulism. These results indicate that grazing can positively affect growing horses in terms of blood components and intestinal microbiota. Moreover, grazing can be helpful to make growing horses healthy through proper exercise.