• Title/Summary/Keyword: environmental DNA

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Spatial Heterogeneity of Bacteria: Evidence from Hot Composts by Culture-independent Analysis

  • Guo, Yan;Zhang, Jinliang;Deng, Changyan;Zhu, Nengwu
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.7
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    • pp.1045-1054
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    • 2012
  • The phylogenetic diversity of the bacteria in hot composting samples collected from three spatial locations was investigated by molecular tools in order to determine the influence of gradient effect on bacterial communities during the thermophilic phase of composting swine manure with rice straw. Total microbial DNA was extracted and bacterial near full-length 16S rRNA genes were subsequently amplified, cloned, restriction fragment length polymorphism-screened and sequenced. The superstratum sample had the highest microbial diversity among the three samples which was possibly related to the surrounding conditions of the sample resulting from the location. The results showed that the sequences related to Bacillus sp. were most common in the composts. In superstratum sample, 45 clones (33%) and 36 clones (27%) were affiliated with the Bacillus sp. and Clostridium sp., respectively; 74 clones (58%) were affiliated with the Clostridium sp. in the middle-level sample; 52 clones (40%) and 29 clones (23%) were affiliated with the Clostridium sp. and Bacillus sp. in substrate sample, respectively. It indicated that the microbial diversity and community in the samples were different for each sampling site, and different locations of the same pile often contained distinct and different microbial communities.

Co-Localization of GABA Shunt Enzymes for the Efficient Production of Gamma-Aminobutyric Acid via GABA Shunt Pathway in Escherichia coli

  • Pham, Van Dung;Somasundaram, Sivachandiran;Park, Si Jae;Lee, Seung Hwan;Hong, Soon Ho
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.710-716
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    • 2016
  • Gamma-aminobutyric acid (GABA) is a non-protein amino acid, which is an important inhibitor of neurotransmission in the human brain. GABA is also used as the precursor of biopolymer Nylon-4 production. In this study, the carbon flux from the tricarboxylic acid cycle was directed to the GABA shunt pathway for the production of GABA from glucose. The GABA shunt enzymes succinate-semialdehyde dehydrogenase (GabD) and GABA aminotransferase (GabT) were co-localized along with the GABA transporter (GadC) by using a synthetic scaffold complex. The co-localized enzyme scaffold complex produced 0.71 g/l of GABA from 10 g/l of glucose. Inactivation of competing metabolic pathways in mutant E. coli strains XBM1 and XBM6 increased GABA production 13% to reach 0.80 g/l GABA by the enzymes co-localized and expressed in the mutant strains. The recombinant E. coli system developed in this study demonstrated the possibility of the pathway of the GABA shunt as a novel GABA production pathway.

3',5'-Cyclic Adenosine Monophosphate (cAMP) as a Signal and a Regulatory Compound in Bacterial Cells (원핵세포에서 신호물질 및 조절인자로서의 3',5'-Cyclic Adenosine Monophosphate의 역할)

  • Chun, Se-Jin;Seok, Young-Jae;Lee, Kyu-Ho
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.289-298
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    • 2006
  • 3',5'-cyclic adenosine monophosphate (cAMP) is an important molecule, which mediates diverse cellular processes. For example, it is involved in regulation of sugar uptake/catabolism, DNA replication, cell division, and motility in various acterial species. In addition, cAMP is one of the critical regulators for syntheses of virulence factors in many pathogenic bacteria. It is believed that cAMP acts as a signal for environmental changes as well as a regulatory factor for gene expressions. Therefore, intracellular concentration of cAMP is finely modulated by according to its rates of synthesis (by adenylate cyclase), excretion, and degradation (by cAMP phosphodiesterase). In the present review, we discuss the bacterial physiological characteristics governed by CAMP and the molecular mechanisms for gene regulation by cAMP. Furthermore, the effect of cAMP on phosphotransferase system is addressed.

Genome Organization and Transcription Response to Harvest of Two Metallothionein-Like Genes in Agaricus bisporus Fruiting Bodies

  • Eastwood, Daniel C.;Bains, Navdeep K.;Henderson, Janey;Burton, Kerry S.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.455-463
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    • 2011
  • Metallothioneins are a class of small cysteine-rich proteins that have been associated with increased tolerance to metal and oxidative stresses in animals, plants, and fungi. We investigated a metallothionein-like (mt-like) gene shown previously to be upregulated in fruiting bodies of the fungus Agaricus bisporus in response to post-harvest storage. Analysis of an A. bisporus genomic DNA cosmid library identified two similar mt-like genes (met1 and met2) arranged as a bidirectional gene pair transcribed from the same promoter region. The promoter contained regulatory elements including 9 metal responsive elements and a CAAT box region 220 bp downstream of met1 that showed striking similarity to a feature in Coprinopsis cinerea mt-like gene promoters. Transcriptional analysis showed that both met genes are significantly and rapidly (within 3 hours) upregulated during post-harvest storage and expression is significantly greater in stipe and cap tissues compared with the gills. However, a strong directionality of the promoter was demonstrated, as transcript levels of met1 were at least two orders of magnitude greater than those of met2 in all samples tested.

Epidemiology of the transmissible gastroenteritis of pigs in Gyeongbuk province (경북지방 돼지에서 발생한TGE의 역학적 특성)

  • 김영환;조광현;김순태;김성국;박인화;손재권
    • Korean Journal of Veterinary Service
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    • v.24 no.3
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    • pp.255-262
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    • 2001
  • Five pigs industry with outbreaks of transmissible gastroenteritis(TGE) in Gyeongbuk province were investigated during the period from January to December 2000. The typical signs of TGE in piglets had transient vomiting and a watery yellowish diarrhea, rapid loss of weight, dehydration and high mortality in pigs under 2 weeks of age. Clinical signs of TGE in growing and finishing pigs and sows were usually limited to inappetence and diarrhea for one or a few days, with vomiting observed in an occasional animal. The detection of TGE viral antigen in epithelial cells of the small intestine had been used in indirect fluorescent antibody test (IFA) for diagnosing TGE in young pigs. WかR had been successfully used to detect the DNA derived from TGEV in specimen of intestinal swabs. Among 5 pigs industry, four showed typical signs of epizootic TGE and one progressing enzootic TGE. It was 22~53 days that the duration of initial clinical disease in TGE outbreaks of pigs investigated in Gyeongbuk province in 2000. However the duration related directly to herd size. Mortality of piglets under 2 weeks of age for duration was 53.2~88.2%, but that of piglets 2~5 weeks of age was 2.5~6.5%. The piglets of 1 weeks of age died mostly during duration of TGE, but varied considerably with husbandry and other environmental factors.

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Use of Terminal Restriction Length Polymorphism (T-RFLP) Analysis to Evaluate Uncultivable Microbial Community Structure of Soil

  • Chauhan, Puneet Singh;Shagol, Charlotte C.;Yim, Woo-Jong;Tipayno, Sherlyn C.;Kim, Chang-Gi;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.127-145
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    • 2011
  • Various environmental ecosystems are valuable sources for microbial ecology studies, and their analyses using recently developed molecular ecological approaches have drawn significant attention within the scientific community. Changes in the microbial community structures due to various anthropogenic activities can be evaluated by various culture-independent methods e.g. ARISA, DGGE, SSCP, T-RFLP, clone library, pyrosequencing, etc. Direct amplification of total community DNA and amplification of most conserved region (16S rRNA) are common initial steps, followed by either fingerprinting or sequencing analysis. Fingerprinting methods are relatively quicker than sequencing analysis in evaluating the changes in the microbial community. Being an efficient, sensitive and time- and cost effective method, T-RFLP is regularly used by many researchers to access the microbial diversity. Among various fingerprinting methods T-RFLP became an important tool in studying the microbial community structure because of its sensitivity and reproducibility. In this present review, we will discuss the important developments in T-RFLP methodology to distinguish the total microbial diversity and community composition in the various ecosystems.

Phenylalanine Ammonia-Lyase Gene (NtPAL4) Induced by Abiotic Stresses in Tobacco (Nicotiana tabacum)

  • Han, Woong;Wang, Myeong-Hyeon
    • Korean Journal of Plant Resources
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    • v.23 no.6
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    • pp.535-540
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    • 2010
  • Phenylalanine ammonia-lyase (PAL), a key enzyme of the phenylpropanoid biosynthesis pathway, is activated by a number of developmental and environmental cues. The coding region of the NtPAL4 gene was 2,154 bp in length, and its deduced protein was composed of 717 amino acids. Sequence analysis of NtPAL4 cDNA from tobacco (Nicotiana tabacum L.) revealed high structural similarity to PAL genes of other plant species. The NtPAL4 gene exists as a single copy in the tobacco plant, and its transcripts were strongly expressed in flowers and leaves. NtPAL4 expression was significantly induced in response to NaCl, mannitol, and cold treatments, but it was not induced by abscisic acid (ABA). NtPAL4 expression decreased gradually after treatment with ABA and $H_2O_2$; however, NtPAL4 transcripts accumulated after treatment with methyl viologen (MV). Our results suggest that the NtPAL4 gene may function in response to abiotic stresses.

DNA Chip Database for the Korean Functional Genomics Project

  • Kim, Sang-Soo
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.10a
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    • pp.11-28
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    • 2001
  • The Korean functional Genomics Project focuses on stomach and liver cancers. Specimens collected by six hospital teams are used in BNA microarray experiments. Experimental conditions, spot measurement data, and the associated clinical information are stored in a relational database. Microarray database schema was developed based on EBI's ArrayExpress. A diagrammatic representation of the schema is used to help navigate over marty tables in the database. Field description, table-to-table relationship, and other database features are also stored in the database and these are used by a PERL interface program to generate web-based input forms on the fly. As such, it is rather simple to modify the database definition and implement controlled vocabularies. This PERL program is a general-purpose utility which can be used for inputting and updating data in relational databases. It supports file upload and user-supplied filters of uploaded data. Joining related tables is implemented using JavaScripts, allowing this step to be deferred to a later stage. This feature alleviates the pain of inputting data into a multi-table database and promotes collaborative data input among several teams. Pathological finding, clinical laboratory parameters, demographical information, and environmental factors are also collected and stored in a separate database. The same PERL program facilitated developing this database and its user-interface.

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Molecular Epidemiology of Cryptococcus neoformans/Cryptococcus gattii Complex Isolates from Pigeon Droppings in Korea

  • Chang, Kyungsoo
    • Biomedical Science Letters
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    • v.19 no.3
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    • pp.213-223
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    • 2013
  • The objectives of this study are to develop a molecular diagnosis to differentiate serotypes and mating-types of C. neoformans/C. gattii complex isolates from pigeon droppings in Korea and to elucidate molecular epidemiology of the isolates. Phenotypes and genotypes of C. neoformans/C. gattii complex isolates were identified by biochemical properties and PCR using specific CNLAC1 gene, respectively. To classify serotypes and mating-types of C. neoformans/C. gattii complex isolates, the five reference strains and thirty-three isolates in Korea were investigated by restriction fragment length polymorphism (RFLP) analysis using CNLAC1 gene for varieties, by random amplified polymorphic DNA (RAPD) for serotyping, and by PCR using specific primer sets for mating typing. All isolates in Korea were belonged to C. neoformans var. grubii (serotype A) by RFLP and RAPD patterns which showed high sensitivity and specificity. Therefore, RFLP and RFLP were available to differentiate varieties and serotypes of C. neoformans. Amplification patterns of the five reference strains by specific PCR for mating typing were differentiable, and all isolates were classified into $MAT{\alpha}$. All C. neoformans environmental isolates in Korea were Cr. neoformans serotype A and $MAT{\alpha}$ which is a more virulent pathogen. This study suggests that RFLP and RAPD are rapid and correct molecular diagnosis tools for epidemiology of C. neoformans/C. gattii complex isolates.

Age-related Increase of Sister Chromatid Exchange Frequency in Bone Marrow Cells of Senescence Accelerated Mouse and Its Inhibition by Chronic Treatment of Ginseng

  • Lim, Heung-Bin;Sohn, Hyung-Ok;Lee, Young-Gu;Kim, Seung-Hyung;Lee, Dong-Wook
    • Toxicological Research
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    • v.11 no.2
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    • pp.261-266
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    • 1995
  • Age-related change in the frequency of spontaneous sister chromatid exchange (SCE) and chromosornal aberrations were investigated in bone marrow cells of accelerated senescence-resistant mice (SAM R1) and senescence accelerated ones (SAM P1). And the effect of chronic treatment of ginseng extract (Panax ginseng C.A. Meyer) on these chromosomal abnormalities was tested in SAM P1. SCE frequency in the cells was progressively increased with age in both mice, but it was consistently higher in SAM P1 than in SAM R1 at all corresponding age. Chromosomal aberrations were, however, not significantly changed with age except that it was slightly increased in only aged SAM P1. Interestingly, the rate of these genetic instabilities in SAM P1 was remarkably retarded by long-term administration of ginseng water extract (0.05% in drinking water). These results suggest that frequency of spontaneous SCE in bone marrow cells increase in parallel with senescence of the mice, and SAM P1 is in the condition of being more exposed than SAM R1 to DNA damaging factors. These also indicate that long-term treatment of ginseng may reduce the genetic damage.

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