• Title/Summary/Keyword: Environmental clone

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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/.

Sequence-Based Screening for Putative Polyketide Synthase Gene-Harboring Clones from a Soil Metagenome Library

  • JI SANG CHUN;KIM DOCKYU;YOON JUNG-HOON;OH TAE-KWANG;LEE CHOONG-HWAN
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.153-157
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    • 2006
  • A soil metagenomic library was constructed using an E. coli-fosmid cloning system with environmental DNAs extracted from Kwangreung forest topsoil. We targeted the genes involved in the biosynthesis of bacterial polyketides. Initially, a total of 36 clone pools (10,800 clones) were explored by the PCR-based method using the metagenomic DNAs from each pool and a degenerate primer set, which has been designed based on the highly conserved regions among ketoacyl synthase (KS) domains in actinomycete type I polyketide synthases (PKS Is). Six clone pools were tentatively selected as positive and further examined through a hybridization-based method for selecting a fosmid clone containing PKS I genes. Colony hybridization was performed against fosmid clones from the 6 positive pools, and finally 4 clones were picked out and confirmed to contain the conserved DNA fragment of KS domains. In this study, we present a simple and feasible sorting method for a desired clone from metagenomic libraries.

Effects of Chemical Carcinogens on the Aldehyde Metabolic Enzymes and Antioxidant Enzymes in Clone 9 Cell (화학적 발암원이 Clone 9 세포의 알데히드대사 효소와 항산화 효소에 미치는 영향)

  • 김현배;염영나;이미영
    • Toxicological Research
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    • v.18 no.3
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    • pp.293-300
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    • 2002
  • Chemical carcinogen-induced alteration of aldehyde metabolic enzymes were examined in clone 9 cell. Diethylnitrosamine (DENA), N-nitrosoethylurea (NEU) and N-nitrosomorpholine (NNM) were wed as model carcinogens. Changes in enzyme activities by repetitive treatment of DENA, NEU or NNM were analyzed in terms of specific activities and activity stainings of the enzymes on the gel. Upon treatment of DENA, lipid peroxide level increased upto 10 fold, indicating strong oxidative stress state of the cell. Notable enhancement of ADH and ALDH activity occurred after DENA treatment, while glutathione-S-transferase activity was slightly increased. Furthermore, about 2.5 fold higher superoxide dismutase (SOD) activity was detected during deactivation of catalase (CAT) activity by repetitive treatment of DENA. However in NEU-treated cell, about 2.3 fold higher ALDH activity was found while ADH activity was slightly increased. Notable increase CAT and SOD could also be found. In contrast, maximum 3.5 fold higher CAT activity occurred during SOD deactivation in NNM-treated cell. These results suggest that there might be different enzymatic responses in relation to cell protection against DENA, NEU or NNM.

Effects of arsenite and variation of microbial community on continuous bio-hydrogen production from molasses using a sequence batch reactor (SBR)

  • William, Dennis Sambai;Lee, Pul-eip;Lee, Tae-jin
    • Environmental Engineering Research
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    • v.20 no.4
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    • pp.370-376
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    • 2015
  • This study investigated the effects of various arsenite concentrations on bio-hydrogen production from molasses using a sequence batch reactor (SBR) operated in a series of three batch cycles. In the first batch cycle, hydrogen production was stimulated at arsenite concentrations lower than 2.0 mg/L, while inhibition occurred at arsenite concentration higher than 2.0 mg/L compared to the control. Hydrogen production decreased substantially during the second batch cycle, while no hydrogen was produced during the third batch cycle at all tested concentrations. The toxic density increased with respect to the increase in arsenite concentrations (6.0 > 1.6 > 1.0 > 0.5 mg/L) and operation cycles (third cycle > second cycle > first cycle). The presence of microorganisms such as Clostridium sp. MSTE9, Uncultured Dysgonomonas sp. clone MEC-4, Pseudomonas parafulva FS04, and Uncultured bacterium clone 584CL3e9 resulted in active stimulation of hydrogen production, however, it was unlikely that Enterobacter sp. sed221 was not related to hydrogen production. The tolerance of arsenite in hydrogen producing microorganisms decreased with the increase in induction time, which resulted in severing the inhibition of continuous hydrogen production.

Physiological Characteristics of Poplar Clones in the Saemanguem Reclaimed Land (새만금간척지에 식재한 포플러 클론의 생리적 특성)

  • Lee, Jong Kyu;Jang, Ji Hwi;Li, Yang;Kim, Hae Naem;Kwak, Myeong Ja;Khaine, Inkyin;Lee, Tae Yoon;Lee, Hyun Kyung;Kim, Ie Rae;Jang, Gyeong Hwan;Lee, Wi Young;Kang, Ho Duck;Woo, Su Young
    • Journal of Korean Society of Forest Science
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    • v.106 no.2
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    • pp.186-195
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    • 2017
  • This research aims to study the physiological characteristics of poplar clones in Saemanguem reclaimed land where has lower minerals and bad drainage than those in paddy field. In July and August, different poplar clones including Populus euramericana clone (Venziano and L.Avanzo) and deltoides hybrid clone (97-18 and 97-19) showed different physiological characteristics. Physiological characteristics of P. euramericana clones were better than those of P. deltoides hybrid clones. Especially, the average net photosynthetic rate ($A_{net}$) of clone Venziano were $18.62{\mu}mol\;CO_2\;m^{-2}{\cdot}s^{-1}$ and $17.11{\mu}mol\;CO_2\;m^{-2}{\cdot}s^{-1}$, respectively. Those were the highest numerical values among other poplar clones. On the other, the total chlorophyll and carotenoid contents of P. deltoides hybrid clones were higher than those of P. euramericana clones in July. Particularly, the total chlorophyll content of clone 97-18 was $2.37mg{\cdot}g^{-1}FW$. In August, those of P. deltoides hybrid clones decreased than in July. In term of the physiological activity such as net photosynthetic rate and stomatal conductance of P. deltoides hybrid clones were much lower in August than in July. As a result, physiological characteristics of P. euramericana Venziano clone was superior than the other clones in Short rotation coppice (SRC) in Saemanguem area.

Cyanobacterial Diversity Analysis Using cpcBA-Intergenic Spacer Region (cpcBA-Intergenic Spacer Region을 이용한 Cyanobacteria의 다양성 분석)

  • Choi Gang-Guk;Park Yong-Ha;Ahn Chi-Yong;Bae Myoung-Sook;Oh Hee-Mock
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.287-292
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    • 2005
  • The cyanobacterial diversity was analyzed by restriction fragment length polymorphism (RFLP) of PCR-amplified rpcBA-Intergenic Spacer (IGS) genes and cpcBA-IGS gene sequencing with a sample collected at Chuso-ri in Daechung Reservoir on March 15, 2005, The Shannon-Weiner diversity index was 0.65, indicating that the cyanobacterial community structure was simple. PCR-RFLP profiles obtained were Phormidium spp. (58 clones), Anabaena spp. (14 clones), Microcystis spp. (4 clones), Spirulina sp. (1 clone) and uncultured cyanobacteria (2 clones). The PCR-RFLP of cpcBA-IGS revealed that Phormidium spp. and Anabaena spp. dominated in the invested sample. As a consequence, it seems that the analysis of functional genes such as cpcBA-IGS can be used for the species identification and community analysis of cyanobacteria.

Analysis on the Microbial Community Structure of Red Granule in the Anaerobic Ammonium Oxidation Reactor (혐기성 암모늄 산화 반응기 내 붉은색 입상슬러지의 미생물 군집구조 분석)

  • Bae, Hyo-Kwan;Park, Kyung-Soon;Chung, Yun-Chul;Jung, Jin-Young
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.10
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    • pp.1055-1064
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    • 2006
  • Extremely slow growing anammox(anaerobic ammonium oxidation) bacteria were cultivated using a combination of UASB(Upflow Anaerobic Sludge Blanket) reactor seeded with anaerobic granular sludge and carbon-fiber cultivating reactor. After 180 days of continuous cultivation, average nitrogen removal rate showed 0.54 kg $N/m^3-day$ when 0.6 kg $N/m^3-day$ of nitrogen loading was applied. The black granule was changed to brown and red granule as continuous operation, and the red granule was highly dependant on the high anammox activity. Microbial community structure of red granule in the UASB reactor was analyzed by molecular methods such as gene cloning, phylogenetic tree analysis, and FISH(Fluorescence In Situ Hybridization) method. As a result of gene cloning and phylogenetic tree analysis, 5 kinds of phylum were found to be Planctomycetes, Proteobacteria, Acidobacteria, Chlorobi and Chloroflexi. 13 clones were matched to anammox bacteria among 51 clones in the red anammox granule. In-silico test which used cloning information and FISH probe of the AMX368 was conducted to detect the presence of anammox bacteria in the red anammox granule. As a result of in-silico test only one clone was exactly matched to AMX368 but 11 clones was mutated one base among 18 bases representing all 12 clones are anammox bacteria. A filamentous Chloroflexi might be related to the granulation of anammox bacteria. As a result of FISH analysis, anammox bacteria was abundant in the red anammox granule.

Evaluation of Microbial PCE Reductive Dechlorination Activity and Microbial Community Structure using PCE-Contaminated Groundwater in Korea (사염화에틸렌(PCE)으로 오염된 국내 4개 지역 지하수 내 생물학적 PCE 탈염소화 활성 및 미생물 군집의 비교)

  • Kim Young;Kim Jin-Wook;Ha Chul-Yoon;Kwon Soo-Yeol;Kim Jung-Kwan;Lee Han-Woong;Ha Joon-Soo;Park Hoo-Won;Ahn Young-Ho;Lee Jin-Woo
    • Journal of Soil and Groundwater Environment
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    • v.10 no.2
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    • pp.52-58
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    • 2005
  • In Korea, little attention has been paid to microbial perchloroethylene (PCE) and/or trichloroethylene (TCE) dechlorination activity and identification of microorganisms involved in PCE reductive dechlorination at a PCE-contaminated aquifer. We performed microcosm tests using the groundwater samples from 4 different contaminated sites (i.e. Changwon A, Changwon B, Bucheon and Yangsan) to assess PCE reductive dechlorination activity. We also adapted molecular techniques to screen what types of known reductive dechlorinators are present at the PCE-contaminated aquifers. In the Changwon A and Changwon B active microcosms where potential electron donors such as sodium propionate, sodium lactate, sodium butyrate, and sodium fumarate, were added, ethylene, an end-product of complete reductive dechlorination of PCE, was detected after a period of 90 days of incubation. In the Bucheon and Yangsan active microcosms, cis-1,2-dichloroethylene (c-DCE) was accumulated without the production of vinyl chloride (VC) and ethylene. Molecular techniques were used to evaluate the microbial community structures in the Changwon B and Yangsan aquifer. We found two sequence types that were closely related to a known PCE to ethylene dechlorinator, named uncultured bacterium clone DCE47, in the Changwon B site clone library. However, in the Yangsan site clone library, no sequence type was closely related to known PCE dechlorinators reported. It is plausible that microorganisms being capable of completely dechlorinating PCE to ethylene may be present in the Changwon B site aquifer. In this study we find that complete PCE reductive dechlorinators are present at some PCE-contaminated sites in Korea. In an engineering point of view this information makes it feasible to apply a biological reductive dechlorination process for remediating PCE- and/or TCE-contaminated aquifers in Korea.