• Title/Summary/Keyword: nucleotide sequence analysis

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Characterization of Phenanthrene Degradation by Sphingomonas sp. HS362 (Sphingomonas sp. HS362에 의한 Phenanthrene 분해특성)

  • Kim Su Hwa;Hong Seung-Bok;Kang Hee Jeong;Ahn Jin-Chul;Jeong Jae Hoon;Son Seung-Yeol
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.201-207
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    • 2005
  • A phenanthrene-degrading bacterium HS362, which is capable of using phenanthrene as a sole carbon and energy source, was isolated from oil contaminated soil. This strain is a gram negative, rod shaped organism that is most closely related to Sphingomonas paucimobilis based on biochemical tests, and belongs to the genus Sphingomonas based on fatty acids analysis. It exhibited more than $99.2{\%}$ nucleotide sequence similarity of 16S rDNA to that of Sphingomonas CF06. Thus, we named this strain as Sphingomonas sp. HS362. It degraded $98{\%}$ of phenanthrene after 10 days of incubation when phenanthrene was added at 500 ppm and $30{\%}$ even when phenanthrene was added at 3000 ppm. Sphingomonas sp. HS362 could also degrade low molecular weight PAHs(Polycyclic aromatic hydrocarbons) such as indole and naphthalene, but was unable to degrade high molecular weight PAHs such as pyrene and fluoranthene. The optimum temperature and pH for phenanthrene degradation were $30^{\circ}C$ and $4{\~}8$, respectively. Sphingomonas sp. HS362 could degrade phenanthrene effectively in the concentration range of NaCl of up to $1{\%}$. Its phenanhrene degrading ability was enhanced by preculture, suggesting the possibility of induction of phenanthrene degrading enzymes. Starch and surfactants such as SDS, Tween 85, and Triton X-100 were also able to enhance phenanthrene degradation by Sphingomonas sp. HS362. It carries five plasmids and one of them, plasmid p4, is considered to be involved in the degradation of phenanthrene according to the plasmid curing experiment by growing at $42^{\circ}C$.

Occurrence of Gray Mold Caused by Botrytis cinerea on Common Fig in Korea (Botrytis cinerea에 의한 무화과 잿빛곰팡이병)

  • Cheong, Seong-Soo;Choi, In-Young;Lee, Wang-Hyu
    • The Korean Journal of Mycology
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    • v.41 no.1
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    • pp.38-41
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    • 2013
  • In 2010 and 2011, gray mold was found on common fig (Ficus carica) fruit grown at the research field of Jeollabuk-do Agricultural Research and Extension Services, Korea. Gray mold symptoms on common fig fruit mainly occurred after harvest season until December. The typical symptom included brown water-soaked rot and fruit decay. The diseased fruit was covered by gray to brown colored conidiophore and conidia. The conidiophores were tree shape and measured $15-33{\times}2{\mu}m$. Conidia on conidiophore were ellipsoidal or lemon shape, colorless, single cell, and measured $7.3-14.6{\times}6.8-11.1{\mu}m$. The nucleotide sequences of the rDNA ITS region obtained from the pure culture of the gray mold on common fig were 100% similar to the sequences of the GenBank accession number HQ171052, EU519210, HQ171053, FN812726, HM849615, and EU563120 of B. cinerea isolates. In phylogenetic tree, the representative isolate was placed within same clade of B. cinerea. Based on the morphological characteristics and analysis of rDNA ITS sequence data, the fungus was identified as B. cinerea.

Genetic Relationships of Coprinus spp. on the Basis of Sequences in ITS II Region (먹물버섯류(Coprinus spp.)의 ITS II 영역 염기서열에 의한 유연관계 분석)

  • Park, Dong-Suk;Go, Seung-Joo;Kim, Yang-Sup;Seok, Soon-Ja;Song, Jae-Kyeong;Yeo, Yun-Soo;Ryu, Jin-Chang;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.27-31
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    • 1999
  • The internal transcribed spacer II regions (ITS II) of the ribosomal DNA gene repeat from Coprinus spp. were amplified using polymerase chain reaction (PCR) and sequenced. Sequences from 11 species including Coprinus comatus, C. atramentarius, C. micaceus, C. lagopus, C. cinereus, C. rhizophorus, C. flocculosus, C. radians, and C. echinosporus were compared. The spacer regions of them were $253{\sim}275$ nucleotide in length and partially contained 5.8S and 25S. The reciprocal homologies of each ITS II sequence among these strains were in the range of $50.6{\sim}100%$. According to the analysis of ITS II sequences, Coprinus spp. were classified into three clusters. Cluster I consisted of Coprinus lagopus, C. cinereus, C. echinosporus, C. rhizophorus, C. niveus, and C. atramentarius. Cluster II comprised C. micaceus, C. flocculosus, C. radians, and C. disseminatus. On the other hand C. comatus is in Cluster III even though this species is belonging to the section Coprinus in morphological aspect. These results suggest that Coprinus comatus, which was considered as a type species of the genus Coprinus in morphological classification, gives a doubt of monophyletic evolution and is assumed to be paraphyletic or polyphyletic.

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Association Analysis of Monocyte Chemotactic Protein-3 (MCP3) Polymorphisms with Asthmatic Phenotypes

  • Park, Byung-Lae;Kim, Lyoung-Hyo;Choi, Yoo-Hyun;Cheong, Hyun-Sub;Park, Hae-Sim;Hong, Soo-Jong;Choi, Byoung-Whui;Lee, June-Hyuk;Uh, Soo-Taek;Park, Choon-Sik;Shin, Hyoung-Doo
    • BMB Reports
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    • v.38 no.1
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    • pp.77-81
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    • 2005
  • The monocyte chemotactic protein-3 (MCP3), on chromosome 17q11.2-q12, is a secreted chemokine, which attracts macrophages during inflammation and metastasis. In an effort to discover additional polymorphism(s) in genes whose variant(s) have been implicated in asthma, we scrutinized the genetic polymorphisms in MCP3 to evaluate it as a potential candidate gene for asthma host genetic study. By direct DNA sequencing in twenty-four individuals, we identified four sequence variants within the 3 kb full genome including 1,000bp promoter region of MCP3; one in promoter region (-420T>C), three in intron (+136C>G, +563C>T, +984G>A) respectively. The frequencies of those four SNPs were 0.020 (-420T>C), 0.038 (+136C>G), 0.080 (+563C>T), 0.035 (+984G>A), respectively, in Korean population (n = 598). Haplotypes, their frequencies and linkage disequilibrium coefficients (|D'|) between SNP pairs were estimated. The associations with the risk of asthma, skin-test reactivity and total serum IgE levels were analyzed. Using statistical analyses for association of MCP3 polymorphisms with asthma development and asthma-related phenotypes, no significant signals were detected. In conclusion, we identified four genetic polymorphisms in the important MCP3 gene, but no significant associations of MCP3 variants with asthma phenotypes were detected. MCP3 variation/haplotype information identified in this study will provide valuable information for future association studies of other allergic diseases.

Lack of Mitochondrial DNA Sequence Divergence between Two Subspecies of the Siberian Weasel from Korea: Mustela sibirica coreanus from the Korean Peninsula and M. s. quelpartis from Jeju Island

  • Koh, Hung-Sun;Jang, Kyung-Hee;Oh, Jang-Geun;Han, Eui-Dong;Jo, Jae-Eun;Ham, Eui-Jeong;Jeong, Seon-Ki;Lee, Jong-Hyek;Kim, Kwang-Seon;Kweon, Gu-Hee;In, Seong-Teak
    • Animal Systematics, Evolution and Diversity
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    • v.28 no.2
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    • pp.133-136
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    • 2012
  • The objective of this study was to determine the degree of mitochondrial DNA (mtDNA) divergence between two subspecies of $Mustela$ $sibirica$ from Korea ($M.$ $s.$ $coreanus$ on the Korean Peninsula and ($M.$ $s.$ $quelpartis$ on Jeju Island) and to examine the taxonomic status of ($M.$ $s.$ $quelpartis$. Thus, we obtained complete sequences of mtDNA cytochrome $b$ gene (1,140 bp) from the two subspecies, and these sequences were compared to a corresponding haplotype of ($M.$ $s.$ $coreanus$, downloaded from GenBank. From this analysis, it was observed that the sequences from monogenic ($M.$ $s.$ $quelpartis$ on Jeju Island were identical to the sequences of four ($M.$ $s.$ $coreanus$from four locations across the Korean Peninsula, and that the two subspecies formed a single clade; the average nucleotide distance between the two subspecies was 0.26% (range, 0.00 to 0.53%). We found that the subspecies $quelpartis$ is not genetically distinct from the subspecies $coreanus$, and that this cytochrome $b$ sequencing result does not support the current classification, distinguishing these two subspecies by pelage color. Further systematic analyses using morphometric characters and other DNA markers are necessary to confirm the taxonomic status of ($M.$ $s.$ $quelpartis$.

A Molecular Sex Identification Using Duplex PCR Method for SRY and ZFX-ZFY Genes in Red Deer and Elk (붉은사슴과 엘크에서 SRY와 ZFX-ZFY 유전자의 Duplex PCR기법을 이용한 성 판별)

  • Han, S.H.;Lee, S.S.;Ko, M.S.;Cho, I.C.
    • Journal of Animal Science and Technology
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    • v.49 no.1
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    • pp.1-8
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    • 2007
  • This study was focused on discriminating the molecular sexes of red deer and elk by duplex polymerase chain reaction(PCR) using two primer sets. Sex differentiation of mammals is primarily dependent on the presence or absence of sex determining region Y(SRY) gene encoded on Y chromosome which plays a key role for male development. Zinc finger X-Y(ZFX-ZFY) gene, one of X-Y homology gene group was found on X- and Y- chromosomes, respectively. At first, the nucleotide sequences were characterized for the intron 9 flanking region of ZFX-ZFY genes. The intron 9 of ZFX and ZFY is 529-bp and 665-bp in length, respectively. A transposable element sequence similar to bovine SINE element Bov-tA was detected only in ZFY gene of Cervidae. Sexing analysis was conducted by duplex PCR assay for amplification of SRY and ZFX-ZFY genes. Two differentially amplified patterns were found: one for females has a common band amplified only from ZFX as a template, and another for males had three bands(a common ZFX and two male-specific ZFY and SRY). On the separate tests using each gene, the results was identical to those from duplex PCR assay. Moreover, the results from PCR assays provide also identical information to phenotypic investigation of individuals of red deer, elk as well as their hybridized progenies collected from two isolated farms. These results suggest that it may be a rapid and precise method for determining the sexes by duplex PCR amplification using Y-chromosome specific SRY and X- and Y- homologous ZFX-ZFY genes showing sexual dimorphism in red deer and elk without any other controls.

Acinetobacter pullorum sp. nov., Isolated from Chicken Meat

  • Elnar, Arxel G.;Kim, Min-Gon;Lee, Ju-Eun;Han, Rae-Hee;Yoon, Sung-Hee;Lee, Gi-Yong;Yang, Soo-Jin;Kim, Geun-Bae
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.526-532
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    • 2020
  • A bacterial strain, designated B301T and isolated from raw chicken meat obtained from a local market in Korea, was characterized and identified using a polyphasic taxonomic approach. Cells were gram-negative, non-motile, obligate-aerobic coccobacilli that were catalase-positive and oxidase-negative. The optimum growth conditions were 30℃, pH 7.0, and 0% NaCl in tryptic soy broth. Colonies were round, convex, smooth, and cream-colored on tryptic soy agar. Strain B301T has a genome size of 3,102,684 bp, with 2,840 protein-coding genes and 102 RNA genes. The 16S rRNA gene analysis revealed that strain B301T belongs to the genus Acinetobacter and shares highest sequence similarity (97.12%) with A. celticus ANC 4603T and A. sichuanensis WCHAc060041T. The average nucleotide identity and digital DNA-DNA hybridization values for closely related species were below the cutoff values for species delineation (95-96% and 70%, respectively). The DNA G+C content of strain B301T was 37.0%. The major respiratory quinone was Q-9, and the cellular fatty acids were primarily summed feature 3 (C16:1 ω6c/C16:1 ω7c), C16:0, and C18:1 ω9c. The major polar lipids were phosphatidylethanolamine, diphosphatidyl-glycerol, phosphatidylglycerol, and phosphatidyl-serine. The antimicrobial resistance profile of strain B301T revealed the absence of antibiotic-resistance genes. Susceptibility to a wide range of antimicrobials, including imipenem, minocycline, ampicillin, and tetracycline, was also observed. The results of the phenotypic, chemotaxonomic, and phylogenetic analyses indicate that strain B301T represents a novel species of the genus Acinetobacter, for which the name Acinetobacter pullorum sp. nov. is proposed. The type strain is B301T (=KACC 21653T = JCM 33942T).

Mitochondrial DNA Analysis of the Fleshy Prawn (Penaeus chinensis) for Stock Discrimination in the Yellow Sea (황해산 대하(Penaeus chinensis)의 계군분석을 위한 미토콘드리아 DNA 분석)

  • HWANG Gyu-Lin;LEE Yong-Chul;CHANG Chung-Soon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.1
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    • pp.88-94
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    • 1997
  • The mitochondrial DNA (mtDNA) restriction fragment length polymorphi는 (RFLPs) of five populations were analyzed to delineate the stocks of Penaeus chinensis (Osbeck) in the Yellow Sea. Comparison of P. chinensis with P. japonicus to clarify the nucleotide divergence between two species was also carried out. Based on the fragment patterns, three composite haplotypes were analyzed in P. chinensis mtDNA as four naplotypes were in P. japonicus. Most individuals of each P. chinensis population are shared by one dominant haplotype. Another two haplotypes haying variations at the C/a I and hull sites were also distributed evenly in the Korean and Chinese populations. It is suggested that the gene exchange occurring between populations in the Yellow Sea is frequent. Average length of the mtDNA molecule was estimated to be about 16.44 kb in P. chinensis and 16.31 kb in P. japonicus, Sequence divergence (p) of mtDNA between two species estimated by using Upholt's (1977) fomula was $13.7\%$.

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Cloning and expression of Streptococcus mutans GS-5 glucosyltransferase (Streptococcus mutans GS-5 Glucosyltransferase의 클로닝과 발현)

  • Kim, Su-Kyeong;Kim, Jae-Gon;Baik, Byeong-Ju;Yang, Yeon-Mi;Lee, Kyung-Yeol;Park, Jeong-Yeol
    • Journal of the korean academy of Pediatric Dentistry
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    • v.35 no.1
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    • pp.73-82
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    • 2008
  • Dental caries is an infectious disease caused by mutans streptococci, and is a primary etiologic agent of dental caries in humans. The molecular pathogenesis of mutans streptococcal-associated dental caries occurs in three phases. Firstly, S. mutans attaches to tooth surface via a cell surface adhesion termed antigen I/II. In the second phase, the glucosyltransferase(GTFs) synthesize polymers like glucans in the presence of sucrose. In the third phase, the multivalent glucans interacts with glucan binding proteins (GBPs) and they make dental plaque and accumulation of microorganisms. Many studies and clinical trials have indicated that a mucosal immune response to these antigens(Ag I/II, GTFs, GBPs) of S. mutans can influence the pathogenesis of dental caries. So these antigens can be important vaccine candidates for immunologic intervention against dental caries. In this study, we cloned the genes for GTFb, GTFc, GTFd from S. mutans GS-5 and did the nucleotide sequence analysis. And the recombinant proteins of GTFd and N-terminus of GTFd were expressed. Intact GTF which we get from this experiment can be used for antibody production specific for any GTF activity domain through animal experiment.

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MLPA Applications in Genetic Testing (유전자진단에 있어서 Multiplex Ligation Dependent Probe Amplification (MLPA)의 이론과 실제)

  • Kim, Gu-Hwan;Lee, Beom-Hee;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.6 no.2
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    • pp.146-154
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    • 2009
  • Multiplex ligation dependent probe amplification (MLPA) is a PCR-based method to detect gene dosage. Since its introduction, MLPA has been used to test a large number of genes for major deletions or duplications. Genetic testing, as a diagnostic tool for genetic disease, has been used primarily to identify point mutations, including base substitutions and small insertions/deletions, using PCR and sequence analysis. However, it is difficult to identify large deletions or duplications using routine PCR- gel based assays, especially in heterozygotes. The MLPA is a more feasible method for identification of gene dosage than another routine PCR-based methods, and better able to detect deleterious deletions or duplications. In addition to detection of gene dosage, MLPA can be applied to identify methylation patterns of target genes, aneuploidy during prenatal diagnoses, and large deletions or duplications that may be associated with various cancers. The MLPA method offers numerous advantages, as it requires only a small amount of template DNA, is applicable to a wide variety of applications, and is high-throughput. On the other hand, this method suffers from disadvantages including the possibility of false positive results affected by template DNA quality, difficulties identifying SNPs located in probe sequences, and analytical complications in quantitative aspects.

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