• 제목/요약/키워드: Denaturing gradient gel electrophoresis (DGGE)

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

  • 박정아;허건영;이정숙;오윤정;김보연;민태익;김치경;안종석
    • 미생물학회지
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    • 제39권1호
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    • pp.45-50
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    • 2003
  • 분자 생물학적 방법 인 DGGE를 이용하여 저온에서 김치가 발효되는 동안 관여하는 미생물의 다양성과 변화양상을 분석하였다. 김치를 저온 ($4^{\circ}C$)에서 발효시키는 60 일 동안 5 일 마다 시료를 채취하였으며, 채취한 김치 시료에서 genomic DNA를 추출하여 실험을 수행하였다. 김치 시료 genomic DNA로부터 16S rDNA의 V3영역을 증폭하여 DGGE를 수행한 결과에서 관찰된 amplicon들의 염기서열을 분석한 결과 저온에서 김치가 발효되는 동안 젖산균들이 주요 미생물 군집으로 나타났으며, 그 중에서도 Weissella koreensis가 발효 전 과정 동안, Lactobacillus sakei의 경우는 발효 10 일째부터, 그리고 Leuconostoc gelidurn은 발효30 일째부터 amplicon들의 농도가 진하게 나타나 이들이 저온에서 김치 발효 과정 동안의 우점종 균주들 임을 알 수 있었다.

유류 분해 근권세균 Rhodococcus sp. 412와 옥수수를 활용한 유류 오염 토양의 정화 (Bioremediation of Oil-Contaminated Soil Using an Oil-Degrading Rhizobacterium Rhodococcus sp.412 and Zea mays.)

  • 홍선화;박혜림;고우리;유재준;조경숙
    • 한국미생물·생명공학회지
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    • 제35권2호
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    • pp.150-157
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    • 2007
  • 디젤 오염 토양 정화를 위해 식물과 미생물의 상호관계를 활용하는 생물복원에 관한 연구를 수행하였다. 디젤을 분해하는 근권 세균인 Rhodococcus sp. 412와 디젤에 내성을 가지고 있는 식물인 옥수수(Zea mays)를 이용하여 디젤로 오염되어진 토양의 디젤 제거능과 미생물 군집변화를 조사하였다. 실험 개시 30일 후, 디젤 오염 토양에서 Rhodococcus sp. 412를 접종한 토양의 옥수수의 성장이 412균주를 접종하지 않은 토양에서의 옥수수 성장보다 약간 우수하였다. 또한 식물을 식재하거나 412균주를 접종한 토양에서 존재하는 토양에서 디젤의 잔류농도도 낮게 나타났다. 이러한 결과를 디젤 오염 토양 정화를 위해 옥수수와 Rhodococcus sp. 412를 동시에 활용하는 것이 유리함을 의미한다. 토양세균 군집 변화를 16S rDNA-PCR과 DGGE(denaturing gradient gel electrophoresis) fingerprinting 방법을 이용하여 분석하였다. 비오염 토양 시료와 디젤 오염토양 시료의 DGGE fingerprint의 유사도는 $20.8{\sim}39.3%$이었다. 또한, 비오염 토양 시료 사이의 DGGE fingerprint의 유사도는 $21.9{\sim}53.6%$, 그리고 디젤 오염 토양 시료 사이의 유사도는 $31.6{\sim}50.0%$이었다. 이러한 결과는 디젤 오염으로 인해 토양 세균 군집구조가 영향을 받았음을 시사한다.

시화호 인공습지의 수질정화 및 암모니아 산화균의 분포 연구 (The Efficacy of Water Purification and Distribution of Ammonia Oxidizing Bacteria in Shihwa Constructed Wetland)

  • 김세윤;김미순;이성희;임미영;이영민;김지열;고광표
    • 한국물환경학회지
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    • 제26권1호
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    • pp.10-18
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    • 2010
  • Water quality and the distribution of ammonia oxidizing bacteria were characterized in constructed wetland of Shihwa lake. Both physico-chemical parameters and fecal indicator microorganisms including total coliforms, E.coli, Enterococcus spp. were measured. In addition, denaturant gradient gel electrophoresis (DGGE) was carried out after PCR amplification of amoA gene from input, output, and wetland sites of the Banwol, Donghwa, and Samhwa stream in Shihwa lake area. Physico-chemical parameters were in proper range for typical nitrifying bacteria to grow and perform their biological activities. Average concentrations of fecal indicator microorganisms of wetland samples were lower than those of input sites. These results suggested that microbial water quality improved by the process of constructed wetland. According to phylogenetic information obtained from DGGE from study sites, distribution of nitrifying bacteria from each of input, output, and wetland were generally distinctive one another. In addition, distribution of nitrifying bacteria between Banwol and Donghwa streams showed higher similarity (52.6%) than this of Samhwa stream (15.2%). These results indicated that characteristics of ammonia oxidizing bacteria in Samhwa were unique in comparison with those of Banwol and Donghwa stream.

Characterization of Microbial Fuel Cells Enriched Using Cr(VI)-Containing Sludge

  • Ryu, Eun-Yeon;Kim, Mi-A;Lee, Sang-Joon
    • Journal of Microbiology and Biotechnology
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    • 제21권2호
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    • pp.187-191
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    • 2011
  • Microbial fuel cells (MFCs) were successfully enriched using sludge contaminated with Cr(VI) and their characteristics were investigated. After enrichment, the charge of the final 10 peaks was 0.51 C ${\pm}$ 1.16%, and the anodic electrode was found to be covered with a biofilm. The enriched MFCs removed 93% of 5 mg/l Cr(VI) and 61% of 25 mg/l Cr(VI). 16S rDNA DGGE profiles from the anodic electrode indicated that ${\beta}$-Proteobacteria, Actinobacteria, and Acinetobacter sp. dominated. This study is the first to report that electrochemically active and Cr(VI)-reducing bacteria could be enriched in the anode compartment of MFCs using Cr(VI)-containing sludge and demonstrates the Cr(VI) removal capability of such MFCs.

혐기성 PCE 탈염소화 관련 미생물 군집 특성

  • 이태호;문부영;박태주
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2004년도 임시총회 및 추계학술발표회
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    • pp.133-137
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    • 2004
  • Tetrachloroethylene(PCE) dechlorination was investigated in an anaerobic enrichment culture from landfill soil. Anaerobic PCE dechlorinating microorganisms could convert 150mg/L of PCE via trichloroethylene(TCE) to cir-1,2-dichloroethylene(CDCE) within 2 days at the optimum temperature of 30 to 35$^{\circ}C$. The enrichment culture could dechlorinate TCE but did not degrade other chlorinated aliphatic compounds, such as cDCE, trans-1,2-dichloroethylene, 1,1-dichloroethylene, 1,1-dichloroethane, 1,2-dichloro- ethane, and 1,1,1-trichloroethane during 5 days incubation. Several isolates from the enrichment culture did not show dechlorinating activity of PCE. Microbial analysis of the dechlorinating enrichment culture by using Polymerase chain reaction-Denaturing gradient gel electrophoresis (PCR-DGGE) method showed that at least three microorganisms were related to the anaerobic PCE dechlorination in the enrichment

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Rapid Identification of Bifidobacteria in Dairy Products by Gene-targeted Species-specific PCR Technique and DGGE

  • Hong, Wei-Shung;Chen, Ming-Ju
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권12호
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    • pp.1887-1894
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    • 2007
  • In this paper, a rapid and reliable gene-targeted species-specific polymerase chain reaction (PCR) technique based on a two-step process was established to identify bifidobacteria in dairy products. The first step was the PCR assay for genus Bifidobacterium with genus specific primers followed by the second step, which identified the species level with species-specific primer mixtures. Ten specific primer pairs, designed from nucleotide sequences of the 16-23S rRNA region, were developed for the Bifidobacterium species including B. angulatum, B. animalis, B. bifidum, B. breve, B. catenulatum, B. infantis, B. longum, B. minimum, B. subtile, and B. thermophilum. This technique was applied to the identification of Bifidobacterium species isolated from 6 probiotic products, and four different Bifidobacterium spp. (B. bifidum, B. longum, B. infantis, and B. breve) were identified. The findings indicated that the 16S-23S rDNA gene-targeted species-specific PCR technique is a simple and reliable method for identification of bifidobacteria in probiotic products. PCR combined with Denaturing Gradient Gel Electrophoresis (DGGE) for identification of the bifidobacteria was also evaluated and compared with the gene-targeted species-specific technique. Results indicated that for fermented milk products consistency was found for both species-specific PCR and PCR-DGGE in detecting species. However, in some lyophilized products, the bands corresponding to these species were not visualized in the DGGE profile but the specific PCR gave a positive result.

PCR-DGGE as a Supplemental Method Verifying Dominance of Culturable Microorganisms from Activated Sludge

  • Zhou, Sheng;Wei, Chaohai;Ke, Lin;Wu, Haizhen
    • Journal of Microbiology and Biotechnology
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    • 제20권11호
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    • pp.1592-1596
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    • 2010
  • To verify the dominance of microorganisms in wastewater biological treatment, PCR-DGGE (denaturing gradient gel electrophoresis) was performed as a supplementary support method for screening of the dominant microorganisms from activated sludge. Results suggest that the dominant microorganisms in activated sludge are primarily responsible for strengthening its effectiveness as a biological treatment system, followed by the non-main dominant microorganisms, whereas the non-dominant microorganisms showed no effects. The degree of microbial abundance present on the profile of PCR-DGGE was in line with the treatment efficiency of augmented activated sludge with isolated cultures, suggesting that PCR-DGGE can be used as an effective supplementary method for verifying culturable dominant microorganisms in activated sludge of coking wastewater.

비점오염물질 처리를 위한 생물막 공정의 운전 및 미생물 군집의 특성 (Study on the Performances and Microbial Community in the Biofilm Process for Treating Nonpoint Source Pollutants)

  • 최기충;박정진;강두기;유재철;변임규;신현석;이태호;박태주
    • 대한환경공학회지
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    • 제30권10호
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    • pp.1021-1027
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    • 2008
  • 본 연구에서는 비점오염물질을 처리하기 위해 생물막 공정이 도입되었다. 반응기내의 생물막의 성장으로 위해 세라믹 담체가 사용되었으며, 담체의 충전률은 각각 5% 및 15(v/v)%였다. 이후, 반응기는 각각 0, 5, 10, 15일의 무강우기간에 따라 회분식으로 운전되었다. COD 및 NH$_4{^+}$-N의 제거효율이 담체 충전률, 온도 및 무강우기간에 따라 조사되었으며, 추가적으로 polymerase chain reaction (PCR)-denaturing gel gradient electrophoresis(DGGE)와 INT-dehydrogenase activity(DHA) test를 통하여 미생물 군집 및 활성도가 해석되었다. 운전 결과, 무강우기간이 늘어남에도 충전률에 관계없이 COD의 제거는 안정적으로 일어났다. COD는 25$^{\circ}C$에서는 6$\sim$8 hr, 10$^{\circ}C$에서는 약 15 hr의 반응시간이 필요하였다. DGGE 분석 결과, 무강우기간이 늘어남에 따라 식종 슬러지에서 발견되는 미생물에서 저니토에서 주로 발견되는 미생물로 변화됨을 확인할 수 있었다. 또한 INT-DHA법에 의한 미생물의 활성도 측정 결과, 15일의 무강우기간에도 활성도의 감소는 관찰되지 않았다.

염도의 변화에 따른 미생물 군집의 변화: 경북 형산강 하류 미생물 군집 변화의 DGGE pattern 분석 (Bacterial Community Structure Shift Driven by Salinity: Analysis of DGGE Band Patterns from Freshwater to Seawater of Hyeongsan River, Korea)

  • 백보람;;황철원;도형기
    • 생명과학회지
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    • 제23권3호
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    • pp.406-414
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    • 2013
  • 강의 하류지역에서 미생물의 군집이 점진적인 염도의 증가에 따라 변한다는 것을 실험적으로 보기 위하여, 경북 형산강의 하류에서부터 연안해역으로 유입되는 곳까지 약 2.91 km 간격으로 0.02%, 1.48%, 2.63%, 3.62%의 염분을 포함하는 물 시료를 얻어 denaturing gradient gel electrophoresis (DGGE)를 수행하였다. 계통분석, 계통수 및 각 시료간의 연관성을 조사한 결과, 미생물 군집의 변화가 염분의 증가에 따라 점진적으로 변화하는 것을 확인하였으며, 이는 염분의 농도가 미생물 군집에 큰 영향을 미치는 요소임을 제시한다. 덧붙여 연안 지역이나 다른 수계환경에 비해 하류지역은 염분의 점진적인 변화로 인해 좁은 면적에 비하여 미생물 다양성이 크고, 이는 곧 특이하고 새로운 종을 찾기에 좋은 장소임을 시사하였다.

Structure and Diversity of Arsenic-Resistant Bacteria in an Old Tin Mine Area of Thailand

  • Jareonmit, Pechrada;Sajjaphan, Kannika;Sadowsky, Michael J.
    • Journal of Microbiology and Biotechnology
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    • 제20권1호
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    • pp.169-178
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    • 2010
  • The microbial community structure in Thailand soils contaminated with low and high levels of arsenic was determined by denaturing gradient gel electrophoresis. Band pattern analysis indicated that the bacterial community was not significantly different in the two soils. Phylogenetic analysis obtained by excising and sequencing six bands indicated that the soils were dominated by Arthrobacter koreensis and $\beta$-Proteobacteria. Two hundred and sixty-two bacterial isolates were obtained from arsenic-contaminated soils. The majority of the As-resistant isolates were Gramnegative bacteria. MIC studies indicated that all of the tested bacteria had greater resistance to arsenate than arsenite. Some strains were capable of growing in medium containing up to 1,500 mg/l arsenite and arsenate. Correlations analysis of resistance patterns of arsenite resistance indicated that the isolated bacteria could be categorized into 13 groups, with a maximum similarity value of 100%. All strains were also evaluated for resistance to eight antibiotics. The antibiotic resistance patterns divided the strains into 100 unique groups, indicating that the strains were very diverse. Isolates from each antibiotic resistance group were characterized in more detail by using the repetitive extragenic palindromic-PCR (rep-PCR) DNA fingerprinting technique with ERIC primers. The PCR products were analyzed by agarose gel electrophoresis. The genetic relatedness of 100 bacterial fingerprints, determined by using the Pearson product-moment similarity coefficient, showed that the isolates could be divided into four clusters, with similarity values ranging from 5-99%. Although many isolates were genetically diverse, others were clonal in nature. Additionally, the arsenic-resistant isolates were examined for the presence of arsenic resistance (ars) genes by using PCR, and 30% of the isolates were found to carry an arsenate reductase encoded by the arsC gene.