• Title/Summary/Keyword: 16S rDNA Denaturing Gradient Gel Electrophoresis

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Analysis of Microbial Communities During Cyanobacterial Bloom in Daechung Reservoir by DGGE (DGGE를 이용한 대청호 수화 발생시기의 세균군집 분석)

  • Ko So-Ra;Park Seong-Joo;Ahn Chi-Yong;Choi Aeran;Lee Jung-Sook;Kim Hee-Sik;Yoon Byung-Dae;Oh Hee-Mock
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.205-210
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    • 2004
  • The change of bacterial communities during cyanobacterial bloom was analyzed by DGGE in Daechung Reservoir from July to October in 2003. The traditional morphological analysis showed that the genera of Microcystis, Chroococcus, Oscillatoria, and Phormidium were dominated. The most frequent band in the DGGE profile by 16S rDNA sequence analysis was identified as Microcystis flos-aquae and the cyanobacterial bloom was peaked on September 2. Oscillatoria spp. were also identified and Aphanizomenon flos-aquae dominated in the middle of August. Judging from the analysis of the digitalized DGGE profiles using the cluster analysis technique, the microbial community on September 2 was considerably different from others. Consequently, it seems that the gene fingerprinting method can give not only the similar results to the traditional morphological method but also additional information on the bacterial species and similarity among the examined microbial communities.

Phylogenetic Diversity of Bacteria Associated with the Marine Sponges, Spirastrella abata and Cinachyrella sp. (해면 Spirastrella abata와 Cinachyrella sp.의 공생 세균의 계통학적 다양성)

  • Cho, Hyun-Hee;Shim, Eun-Jung;Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.177-182
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    • 2010
  • The bacterial community structure of two marine sponges, Spirastrella abata and Cinachyrella sp. collected from Jeju Island, in April 2009, was analyzed by 16S rDNA-denaturing gradient gel electrophoresis (DGGE). DGGE banding patterns indicated 8 and 7 bands for Spirastrella abata and Cinachyrella sp., respectively. Comparative sequence analysis of variable DGGE bands revealed from 92% to 100% similarity to the known published sequences. The bacterial groups associated with Spirastrella abata were Alphaproteobacteria and Deltaproteobacteria. The bacterial community of Cinachyrella sp. consisted of Alphaproteobacteria, Gammaproteobacteria, and Actinobacteria. Alphaproteobacteria was common and predominant in both the sponge species. Deltaproteobacteria was found only in Spirastrella abata while Actinobacteria and Gammaproteobacteria were found only in Cinachyrella sp. The results revealed that though the common bacterial group was found in both the sponges, the bacterial community profiles differed between the two sponge species obtained from the same geographical location.

A Method for Comparing Multiple Bacterial Community Structures from 16S rDNA Clone Library Sequences

  • Hur, Inae;Chun, Jongsik
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.9-13
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    • 2004
  • Culture-independent approaches, based on 16S rDNA sequences, are extensively used in modern microbial ecology. Sequencing of the clone library generated from environmental DNA has advantages over fingerprint-based methods, such as denaturing gradient gel electrophoresis, as it provides precise identification and quantification of the phylotypes present in samples. However, to date, no method exists for comparing multiple bacterial community structures using clone library sequences. In this study, an automated method to achieve this has been developed, by applying pair wise alignment, hierarchical clustering and principle component analysis. The method has been demonstrated to be successful in comparing samples from various environments. The program, named CommCluster, was written in JAVA, and is now freely available, at http://chunlab.snu.ac.kr/commcluster/.

Change of Phytoplankton Community by Ultrasonication in Eutrophic Ponds (부영양 연못에서 초음파 작동에 따른 식물플랑크톤의 군집 변화)

  • Ko, So-Ra;Ahn, Chi-Yong;Joung, Seung-Hyun;Kim, Hee-Sik;Oh, Hee-Mock
    • Korean Journal of Environmental Biology
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    • v.24 no.3
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    • pp.221-229
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    • 2006
  • The effects of ultrasonication on phytoplankton were investigated in two ponds in which physicochemical and biological water quality was similar, one as a treatment and the other as a control. The samples were collected from August 18 to September 30 in 2003. Traditional morphological analysis showed that Bacillariophyceae dominated phytoplankton community in both ponds. The abundance of Cyanophyceae was lower in the phytoplankton community of the sonicated pond than that of control pond. We used DGGE (denaturing gradient gel electrophoresis) to analyze the diversity and change of phytoplankton community in two ponds. The DGGE banding patterns of 16S rRNA gene and sequence analysis demonstrated that Oscillatoria acuminata and CFB (Cytophaga-Flavobacterium-Bacteroides) group bacterium appeared in the treated pond, and the control pond was dominated by Synechococcus sp. and Aphanizomenon flos-aquae. Especially, Pseudanabaena sp. dominated during the ultrasonic cessation in the treated pond. The DGGE profiles of 18S rRNA gene and sequence analysis showed that the treated pond was dominated by Chlamydomonas reinhardtii and the control pond by C. reinhardtii and Pteromonas protracta. In conclusion, the ultrasonication affected the reduced growth of cyanobacteria, particularly Pseudanabaena.

Comparison of Microbial Diversity of Korean Commercial Makgeolli Showing High ${\beta}$-Glucan Content and High Antihypertensive Activity, Respectively

  • Min, Jin-Hong;Kim, Young-Hun;Kim, Jae-Ho;Choi, Shin-Yang;Lee, Jong-Soo;Kim, Ha-Kun
    • Mycobiology
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    • v.40 no.2
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    • pp.138-141
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    • 2012
  • We measured physiological functionalities, including antihypertensive angiotensin I-converting enzyme inhibitory activity and immun-stimulating ${\beta}$-glucan content for sixty kinds of Makgeolli that is commercially available from the market. As a result, we selected R-12 commercial raw Makgeolli, with a high content of immuno-stimulating ${\beta}$-glucan, and R-14 commercial raw Makgeolli, exhibiting high antihypertensive activity. Due to the similarities in their overall physicochemical properties and raw materials used for fermentation, we compared the microbial flora in order to investigate the reason for the differences in their functionalities. Nested PCR and denaturing gradient gel electrophoresis for yeasts and bacteria were performed for analysis of microbial diversity of two different kinds of Makgeolli (i.e., R-12, R-14), which showed immuno-stimulating ${\beta}$-glucan content and exhibited a very high level of antihypertensive activity, respectively. Analysis of the 18S rDNA amplicon revealed a major presence of the yeast strain Pichia burtonii in every Makgeolli sample. Analysis of the 16S rDNA amplicon revealed a predominance of lactic acid bacteria, and the most frequent lactic acid bacteria were Lactobacillus ingluviei, L. fermentum, and L. harbinensis, and Lactobacillus sp. Among these, L. harbinensis was detected only in R-12 and L. ingluviei was found only in R-14. Different functionalities from the individual commercially available Makgeolli may be attributed to actions of different microbial flora during fermentation.

Analysis of Microbial Community During the Anaerobic Dechlorination of Perchloroethylene and Trichloroethylene (Perchloroethylene과 Trichloroethylene의 혐기적 탈염소화 및 미생물 군집 분석)

  • Lee Jae-Won;Kim Byung-Hyuk;Ahn Chi-Yong;Kim Hee-Sik;Yoon Byung-Dae;Oh Hee-Mock
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.281-286
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    • 2005
  • In this study, the anaerobic enrichment cultivation was performed with the sediments and the dredged soils from the cities of Ulsan, Masan, Yeosu, Gwangyang, Ansan and Seongnam. Acetate as an electron donor and PCE (perchloroethylene) or TCE (trichloroethylene) as an electron acceptor were injected into the serum bottle with an anaerobic medium. After the incubation of 12 weeks, the removal efficiency of PCE was highest at $70\%$ in the treatment with the sediment of Ulsan. Also, the bacterial community structure was analyzed by D-DGGE (double denatured gradient gel electrophoresis) through PCR-based 16S rDNA approaches. The dominant species id the anaerobic enrichment were found to belong to the genus of Desulfovibrio.

Application of Denaturing Gradient Gel Electrophoresis to Estimate the Diversity of Commensal Thermophiles

  • Bae, Jin-Woo;Kim, Joong-Jae;Jeon, Che-Ok;Kim, Kwang;Song, Jae-Jun;Lee, Seung-Goo;Poo, Har-Young;Jung, Chang-Min;Park, Yong-Ha;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.1008-1012
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    • 2003
  • Symbiobacterium toebii has been reported as a thermophile exhibiting a commensal interaction with Geobacillus toebii. The distribution of the commensal thermophiles in various soils was investigated using a denaturing gradient gel electrophoresis (DGGE). Based on the DGGE analysis, the enrichment condition for the growth of Symbiobacterium sp. was found to also enrich populations of several other microbial spp. as well as Symbiobacterium sp. In the enrichment experiment, several different 16S rDNA sequences of commensal thermophiles were detected in all of the soil samples tested, indicating that commensal thermophiles are widely distributed in various soils.

Change of Microbial Communities in Kimchi Fermentation at Low Temperature (김치의 저온 발효 중 미생물 변화 양상)

  • 박정아;허건영;이정숙;오윤정;김보연;민태익;김치경;안종석
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.45-50
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    • 2003
  • The diversity and change of microbial communities during kimchi fermentation at $4^{\circ}C$ were analyzed by denaturing gradient gel electrophoresis (DGGE). Kimchi samples were taken every 5 days over the fermentation periods (for 60 days) to extract total DNA for DGGE analysis. Touchdown polymerase chain reaction was performed to amplify the V3 region of 16S rRNA gene. Sequencing results of partial 16S rDNA amplicons from DGGE profiles revealed that lactic acid bacteria (LAB), especially Weissella koreensis, Lactobacillus sakei and Leuconostoc gelidum were dominants in kimchi fermentation at $4^{\circ}C$. And we knew that W. koreensis steadily existed throughout the whole fermentation period, also Lb. sakei and Leuc. gelidum appeared from 10th day and 30th day of fermentation time, respectively and then these species were to be dominant microorganisms.

Detection of Nocardia sp. Hl7-1 by PCR during Bioremediation of Crude Oil-Contaminated Soil (원유 오염토양의 Bioremediation과정 동안 PCR을 이용한 Nocardia sp. Hl7-1의 검출)

  • Baek, Kyung-Hwa;Lee, Young-Ki;Lee, In-Sook;Oh, Hee-Mock;Yoon, Byung-Dae;Kim, Hee-Sik
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.91-95
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    • 2004
  • For the detection of the oil-degrading bacterium, Nocardia sp. Hl7-1, inoculated during the bioremediation of oil-contaminated soil, a species-specific primer was constructed based on the 16S rDNA sequence of this strain. Two forward primers and two reverse primers were designed and tested against both closely and distantly related bacterial strains. All the primers designed were specific to the Nocardia sp. H17-1. Particularly, primer sets NH169F-NH972R and NH575F-NH972R could be used to detect 50 fg of template DNA and TEX>$1.2${\times}$10^4$ CFU/g of sandy soil. These two PCR primer sets successfully detected the H 17-1 strain in the oil-con-laminated soil samples containing heterogeneous DNA. We also conformed the primer specificity by restriction-enzyme cleavage of the PCR products and denaturing gradient gel electrophoresis.

Microbial Community Profiling in cis- and trans-Dichloroethene Enrichment Systems Using Denaturing Gradient Gel Electrophoresis

  • Olaniran, Ademola O.;Stafford, William H.L.;Cowan, Don A.;Pillay, Dorsamy;Pillay, Balakrishna
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.560-570
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    • 2007
  • The effective and accurate assessment of the total microbial community diversity is one of the primary challenges in modem microbial ecology, especially for the detection and characterization of unculturable populations and populations with a low abundance. Accordingly, this study was undertaken to investigate the diversity of the microbial community during the biodegradation of cis- and trans-dichloroethenes in soil and wastewater enrichment cultures. Community profiling using PCR targeting the l6S rRNA gene and denaturing gradient gel electrophoresis (PCR-DGGE) revealed an alteration in the bacterial community profiles with time. Exposure to cis- and trans-dichloroethenes led to the disappearance of certain genospecies that were initially observed in the untreated samples. A cluster analysis of the bacterial DGGE community profiles at various sampling times during the degradation process indicated that the community profile became stable after day 10 of the enrichment. DNA sequencing and phylogenetic analysis of selected DGGE bands revealed that the genera Acinetobacter, Pseudomonas, Bacillus, Comamonas, and Arthrobacter, plus several other important uncultured bacterial phylotypes, dominated the enrichment cultures. Thus, the identified dominant phylotypes may play an important role in the degradation of cis- and trans-dichloroethenes.