• Title/Summary/Keyword: l6S rDNA

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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
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    • v.55 no.4
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    • pp.368-375
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    • 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.

Gene Expression of Enzymes Related to Glutathione Metabolism in Anticancer Drug-resistant L1210 Sublines (항암제 내성 L1210세포의 Glutathione 대사 관련효소 유전자의 발현 양상)

  • Kim, Seong-Yong;Kim, Jae-Ryong;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.12 no.1
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    • pp.32-47
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    • 1995
  • Glutathione(GSH) has a very important role in detoxification of cells and is closely related to antitumor drug-resistance of cancer cells. In order to evaluate the importance of glutathione metabolism in the drug-resistant cancer cells, the concentration of celluar GSH and activities of ${\gamma}$-glutamylcysteine synthetase(GCS), ${\gamma}$-glutamyl transpeptidase (GGT) and glutathione S-transferases(GST) in the adriamycin, vincristine, or cisplatin resistant L1210 (L1210AdR, L1210VcR, or L1210Cis) sublines were measured. Expression and amplification of GCS, GGT, and GST-${\pi}$ genes were also observed in the parent Ll210 and the drug-resistant Ll210 sublines. The concentration of GSH was increased 5.34 fold in L1210Cis, 2.83 fold in L1210VcR, and 1.78 fold in L1210AdR, compared to L1210. The activities of GCS and GGT were increased in drug-resistant L1210 sublines. The GST activity was increased in L1210VcR and L1210Cis but decreased in L1210AdR compared to Ll210. Expression of GCS, GGT, and GST-${\pi}$ genes were increased in the resistant L1210 sublines compare to the parent L1210 in northern blot analyses. Overexpression of GCS, GGT, and GST-${\pi}$ were observed in the resistant sublines, and the increases of the concentration of glutathione and the activities of GCS and GGT in the resistant sublines may be involved in a part of the drug-resistance in the resistant sublines.

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Effects of Electrochemical Reduction Reactions on the Biodegradation of Recalcitrant Organic Compounds (ROCs) and Bacterial Community Diversity

  • Lee, Woo-Jin;Lee, Jong-Kwang;Chung, Jin-Wook;Cho, Yong-Ju;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1230-1239
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    • 2010
  • Five bacterial species, capable of degrading the recalcitrant organic compounds (ROCs) diethyleneglycol monomethylether (DGMME), 1-amino-2-propanol (APOL), 1-methyl-2-pyrrolidinone (NMP), diethyleneglycol monoethylether (DGMEE), tetraethyleneglycol (TEG), and tetrahydrothiophene 1,1-dioxide (sulfolane), were isolated from an enrichment culture. Cupriavidus sp. catabolized $93.5{\pm}1.7$ mg/l of TEG, $99.3{\pm}1.2$ mg/l of DGMME, $96.1{\pm}1.6$ mg/l of APOL, and $99.5{\pm}0.5$ mg/l of NMP in 3 days. Acineobacter sp. catabolized 100 mg/l of DGMME, $99.9{\pm}0.1$ mg/l of NMP, and 100 mg/l of DGMEE in 3 days. Pseudomonas sp.3 catabolized $95.7{\pm}1.2$ mg/l of APOL and $99.8{\pm}0.3$ mg/l of NMP. Paracoccus sp. catabolized $98.3{\pm}0.6$ mg/l of DGMME and $98.3{\pm}1.0$ mg/l of DGMEE in 3 days. A maximum $43{\pm}2.0$ mg/l of sulfolane was catabolized by Paracoccus sp. in 3 days. When a mixed culture composed of the five bacterial species was applied to real wastewater containing DGMME, APOL, NMP, DGMEE, or TEG, 92~99% of each individual ROC was catabolized within 3 days. However, at least 9 days were required for the complete mineralization of sulfolane. Bacterial community diversity, analyzed on the basis of the TGGE pattern of 16S rDNA extracted from viable cells, was found to be significantly reduced in a conventional bioreactor after 6 days of incubation. However, biodiversity was maintained after 12 days of incubation in an electrochemical bioreactor. In conclusion, the electrochemical reduction reaction enhanced the diversity of the bacterial community and actively catabolized sulfolane.

Statistical Optimization for Production of Carboxymethylcellulase from Rice Hulls by a Newly Isolated Marine Microorganism Bacillus licheniformis LBH-52 Using Response Surface Method (통계학적인 방법과 왕겨를 기질로 사용하여 해양에서 분리한 Bacillus licheniformis LBH-52 를 사용한 carboxymethylcellualse의 생산조건 최적화)

  • Kim, Hye-Jin;Gao, Wa;Chung, Chung-Han;Lee, Jin-Woo
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1083-1093
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    • 2011
  • A microorganism utilizing rice hulls as a substrate for the production of carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus lincheniformis by analyses of its 16S rDNA sequences. The optimal carbon and nitrogen sources for production of CMCase were found to be rice hulls and ammonium nitrate. The optimal conditions for cell growth and the production of CMCase by B. lincheniformis LBH-52 were investigated using the response surface method (RSM). The analysis of variance (ANOVA) of results from central composite design (CCD) indicated that a highly significant factor ("probe>F" less than 0.0001) for cell growth was rice hulls, whereas those for production of CMCase were rice hulls and initial pH of the medium. The optimal conditions of rice hulls, ammonium nitrate, initial pH, and temperature for cell growth extracted by Design Expert Software were 48.7 g/l, 1.8 g/l, 6.6, and 35.7$^{\circ}C$, respectively, whereas those for the production of CMCase were 43.2 g/l, 1.1 g/l, 6.8, and 35.7$^{\circ}C$. The maximal production of CMCase by B. lincheniformis LBH-52 from rice hulls under optimized conditions was 79.6 U/ml in a 7 l bioreactor. In this study, rice hulls and ammonium nitrate were developed to be substrates for the production of CMCase by a newly isolated marine microorganism, and the time for production of CMCase was reduced to 3 days using a bacterial strain with submerged fermentation.

Genetic Variation and Population Specific Mitochondrial DNA Haplotype Found in the Jeju Native Pig Population (제주재래돼지 집단서 집단특이적 mtDNA Haplotype과 유전적 다양성)

  • Han, S.H.;Cho, I.C.;Lee, C.E.;Lee, S.S.;Kang, S.Y.;Choi, Y.L.;Oh, W.Y.;Sung, P.N.;Ko, S.B.;Oh, M.Y.;Ko, M.S.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.917-924
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    • 2004
  • Using PCR-RFLP haplotyping for the mitochondrial DNA(mtDNA) fragment containing the NADH dehydrogenase 2 gene(ND2) and three tRNA genes(tRNA-Met, tRNA-Trp and tRNA-Ala), we characterized the genetic diversity of five pig breeds including Jeju native pigs. mtDNA polymorphisms showing distinct cleavage patterns were found in the pig breeds. Two digestion patterns were detected when HaeIII- and Hinfl-RFLP, and four in the Tsp5091-RFLP analyses. Combining the three restriction enzyme digestion patterns found in five different pig breeds, four mtDNA haplotypes were observed and the haplotype frequencies were significantly different by the pig breeds. A monomorphic haplotype, mtWB, was observed in both Korean wild boars and Large White pigs. Both Duroc and Landrace pigs contained two haplotypes suggesting their multiple maternal lineages. Jeju native pig has two haplotypes(mtJN and mtJD). Of these, mtJN is identified as a Jeju native pig specific haplotype. This study suggested that more than two progenitor populations have been taken part in the domestication process of the Jeju native pig population, and/or probably subsequent crossing with other pig breeds from near east Asia. Unlike with our prediction, there was no direct evidence under molecular levels on the maternal introgression of Korean wild boar in the domestication of Jeju native pigs. In conclusion, specificity of mtDNA haplotypes related to pig breeds win be useful for identifying the maternal lineage as wen as constructing the genealogical pedigree in pigs.

Use of 16S-23S rRNA Intergenic Spacer Region for Rapid Detection of Vibrio fluvialis (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio fluvialis의 검출)

  • 강현실;허문수;이제희
    • Korean Journal of Environmental Biology
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    • v.21 no.1
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    • pp.77-85
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    • 2003
  • We have examined the 16S-23S rRNA intergenic spacer regions (ISR) of Vibrio fluvialis. ISRs were PCR amplified, cloned into a plasmid vector and then sequenced. As results of ISR nucleotide sequence analysis, total of 6 clones were isolated depending on the size. The clones were different in both the number and the composition of the tRNA genes, and were designated ISR-A, ISR-E, ISR-El, ISR-lA, ISR-EKV, ISR-EKAV. ISR-A contains $tRNA^{Ala}$; ISR-lA, $tRNA^{Ile}$-$tRNA^{Ala}$; ISR-EKV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Val}$;ISE-EKAV, $tRNA^{GIu}$-$tRNA^{Lys}$-$tRNA^{Ala}$-$tRNA^{Val}$; ISR -E and E1, $tRNA^{GIu}$ clusters. ISR-EKV was shown to be a minor type out of the six ISR types and showed a very limited homology between ISR-EKV from V, fluvialis and ISRa from other Vibrio species. Therefore ISR-EKV sequence was used to design species-specific primers to detect V, fiuvialis from other Vibrio species by PCR reaction. The specificity of the primers was examined using genomic DNA of other Vibrios as templates for PCR reaction. The result showed that PCR can be a useful method to detect V. fluvialis among Vibrio species in a single PCR reaction.

Subgenus classification of Accnthcmoebc by riboprinting (Riboprinting에 의한 가시아메바속의 분류)

  • 정동일;유학선
    • Parasites, Hosts and Diseases
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    • v.36 no.2
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    • pp.69-80
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    • 1998
  • Subgenus classification of Acanthcmoeba remains uncertain. Twenty-three reference strains of Acanthnmoeba including 18 (neo)type-strains were subjected for classification at the subgenus level by riboprinting, PCR/RFLP analysis of 185 rRNA gene (rDNA) . On the dendrogram reconstructed on the basis of riboprint analyses, two type- strains (A. astronwxis and A. tubinshi) of morphological group 1 diverged early from the other strains and were quite distinct from each other. Four type-strains of morphological group 3. A. culbertsoni, A. polestinensis, A. healyi were considered taxonomically valid, but A. pustulosn was regarded as an invalid synonym of A. pclestinensis. Strains of morphological group 2 were classified into 6 subgroups. Among them, A. giulni which has an intron in its 185 rDNA was the most divergent from the remaining strains. Acanthcmoebc ccstellanii Castellani, A. quinc Vil3, A. Iugdunensis L3a. A. poIyphage Jones, A. trinngularis SH621, and A. cqstellanii Ma strains belonged to a subgroup, A. castellanii complex. However, A. quinc and A. Lugnunensis were regarded as synonyms of A. ccstellanii. The Chang strain could be regarded as A. hatchetti. Acanthcmoebo nauritaniensis, A. niuionensis, A. paranivionensis could be considered as synonyms of A. rhwsodes. Neff strain was regarded as A. polyphage rather than as A. castellanii. It is likely that riboprinting can be applied for rapid identification of Acnnthcmoebc isolated from the clinical specimens and environments.

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Chemosystematics and Molecular Phylogeny of a New Bioflocculant-Producing Aspergillus Strain Isolated from Korean Soil

  • Kim, Gi-Young;Ha, Myoung-Gyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.870-872
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    • 1999
  • The ubiquinone and G+C contents of the bioflocculant-producing fungus, a new Aspergillus strain, were detennined using high-perfonnance liquid chromatography. The internal transcribed spacers 1 and 2 (ITS1 and ITS2), and the 5.8S ribosomal DNA (rDNA) of the strain were amplified and sequenced. The strain contained ubiquinone-l0($H_2$)as a major quinone and the G+C content was 49 mol%. A phylogenetic analysis of the ITS regions indicated that the strain belonged to the genus Aspergillus according to its previously classified morphological characteristics. Based on a sequence homology search, the strain was most closely related to Petromyces muricatus (anamorph, A. muricatus; accession number, AJ005674). The sequence of a new Aspergillus strain in ITS1 and ITS2, and 5.8S rDNA showed 97% homology to P. muricatus. Therefore, the strain is believed to be a new bioflocculant-producing Aspergillus strain.

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Characteristics and Frequencies of Alternative Initiation Codon(AIC) of mtDNA ND2 in Five Pig Breeds (돼지 5품종에 있어서 mtDNA ND2 유전자의 선택적 개시코돈의 특성과 빈도)

  • Han, S.H.;Cho, I.C;Choi, Y.L.;Lee, C.E.;Ko, M,S.;Kim, J.H.;Seo, B.Y.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.903-908
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    • 2004
  • Alternative initiation codon(AlC) has been reported in the mitochondrial genes in various mammalian species. We investigated the AlC of mitochondrial NADH dehydrogenase 2 gene(rntDNA ND2) in five pig breeds. Two kinds of initiation codons(ATA/ATf) showing different frequencies among tested pig breeds were used. While all Large White pigs had ATA as an initiator methionine codon, all Landrace pigs had ATf. The other breeds(Berkshire, Duroc and Hampshire) had both initiation codons with the ATA frequencies, 91.9, 21.3 and 60.00/0, respectively. In the previous reports, all Chinese indigenous pig breeds were identified to have unique initiation codon ATA. Although the effect of Ale on the translation of mtDNA ND2 has not been studied in this study, AlC patterns in mtDNA ND2 will contribute to the maternity test using molecular markers in pig breeding.

The Biological Treatment of Soil Washing Water Contaminated with Heavy Metal (중금속오염 토양 세척수의 생물학적 처리)

  • Jeong, Jeong-Hwa;Seo, Pil-Soo;Kong, Sung-Ho;Seo, Seung-Won;Kim, Min-Kyoung;Lee, Jong-Yeol;Lee, Sang-Seob
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.11
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    • pp.1222-1227
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    • 2006
  • In this study, nine strains were isolated from heavy metal-contaminated soil in a mine. The high efficiency bacteria, JH1, to be able removal cadmium and copper, was selected by the screen test. JH1 was identified as Ralstonia eutropha by 16S rDNA analysis, fatty acid analysis, and its morphological and biochemical characteristics. After the cadmium-contaminated soil was washed with citric acid solution(pH 6, 10 mM), Ralstonia eutropha JH1 was inoculated in the soil washing water. In order to determine the optimal cell concentration for inoculation, cell concentrations were considered in 0.5, 1.0, 2.0, 4.0 g/L, respectively. The removal efficiencies for cadmium in each cell concentration of Ralstonia eutropha JH1 were 49.9, 84.4, 89.7% and 89.9% of 110 mg/L(Cd), after 5 days culture in soil washing water. When Ralstonia eutropha JH1 was inoculated in soil washing water containing each cadmium(110 mg/L) and copper(100 mg/L), each of them was removed completely during 6 days culture. The completely removing time for cadmium and copper in each low concentration, 10, 30 and 60 mg/L were 12, 18 and 48 hrs, respectively.