• Title/Summary/Keyword: Phylogenetic

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Prevalence and Genotypes of Transfusion-Transmitted Virus in Children with Hepatitis and Normal Control (정상 소아와 간염 환자에서 Transfusion-Transmitted Virus의 감염상태와 유전자형)

  • Chung, Ju Young;Han, Tae Hee;Hwang, Eung Soo;Ko, Jae Sung;Seo, Jeong Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.8 no.2
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    • pp.202-212
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    • 2005
  • Purpose: Transfusion transmitted virus (TTV) is a newly discovered virus and to date the contribution of TTV to liver disease remains unclear. Little is known about the frequency of TTV infection in children in Korea. The purpose of this study was to investigate the prevalence and genotypic distribution of TTV carried by healthy children and patients with hepatitis in Korea. Methods: Eighty eight of healthy children and three groups of patients with hepatitis-14 patients with chronic hepatitis B, 12 patients with chronic hepatitis C and 25 patients with hepatitis of unknown etiology-were tested. TTV DNA was detected by semi-nested PCR using primer sets generated from N-22 region and from 5' noncoding region (NCR) of the viral genome. PCR products derived from 8 patients with hepatitis and from 11 healthy children were sequenced and a phylogenetic tree was constructed. Results: TTV was found by PCR with N22 primer in 11.3% of healthy children, 28.5% of children with hepatitis B, 25% of children with hepatitis C, 24% of children with hepatitis of unknown etiology. TTV DNA was found by PCR with 5'NCR primer in 32.9% of healthy children, 71.4% of patients with chronic hepatitis B, in 50% of patients with hepatitis C and in 48% of patients with hepatitis of unknown etiology. TLMV DNA was found in 48.9% of healthy children, 21.4% of patients with hepatitis B, 16.6% of patients with hepatitis C, 40% of patients with hepatitis of unknown etiology. Among the sequenced isolates, 10(52%) belonged to genotype 1 (G1) and others belonged to genotype 2 (G2) or genotype 3 (G3). Among the G1 sequences, 7 were grouped as G1a. Conclusion: TTV infection was common in healthy children and in patients with hepatitis. But, the prevalence of TTV DNA by 5'NCR primer was relatively high in patients with hepatitis B and there may be some association between TTV and hepatitis B virus infection. G1 was the major genotype of the studied population.

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Diversity and Characteristics of Rhizosphere Microorganisms Isolated from the Soil around the Roots of Three Plants Native to the Dokdo Islands (독도의 자생식물의 근권에서 분리한 원핵 미생물의 다양성 분석)

  • Kim, Ye-Eun;Yoon, Hyeokjun;You, Young-Hyun;Kim, Hyun;Seo, Yeonggyo;Kim, Miae;Woo, Ju-Ri;Nam, Yoon-Jong;Irina, Khalmuratova;Lee, Gyeong-Min;Song, Jin-Ha;Jin, Young-Ju;Kim, Jong-Guk;Seu, Young-Bae
    • Journal of Life Science
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    • v.24 no.4
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    • pp.461-466
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    • 2014
  • Three plant species, Aster sphathulifolius, Sedum oryzifolium, and Lysimachia mauritiana, native to the Dokdo Islands in South Korea, were examined for rhizosphere microorganisms by using 16S rDNA sequences. Nine species of rhizosphere microorganisms were isolated from the three native plant species, respectively. Phylogenetic analysis showed that the microorganisms could be classified into 19 species belonging to four phyla (Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria), and the characteristics of the microbes were confirmed. Rhizosphere microorganisms from the six orders (Bacillales, Corynebacteriales, Flavobacteriales, Micrococcales, Oceanospirillales, and Rhodobacterales) were isolated from S. oryzifolium. From L. mauritiana, microbes belonging to the seven orders (Bacillales, Flavobacteriales, Micrococcales, Oceanospirillales, Rhizobiales, and Rhodobacterales) were isolated. From A. sphathulifolius, the six orders of rhizosphere microorganisms (Alteromonadales, Bacillales, Corynebacteriales, Flavobacteriales, Micrococcales, and Rhizobiales) were isolated. These data showed that Actinobacteria and Proteobacteria were the dominant phyla for the rhizosphere of all three plants. To confirm the bacterial diversity in rhizospheres, Shannon's diversity index (H') was used at the genus level. In these data, the rhizosphere from S. oryzifolium and L. mauritiana had more diverse bacteria compared to that from A. sphathulifolius.

Paenibacillus kimchicus sp. nov., an antimicrobial bacterium isolated from Kimchi (김치로부터 분리된 항균 활성 세균 Paenibacillus kimchicus sp. nov.)

  • Park, A-rum;Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.319-326
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    • 2016
  • An antimicrobial bacterium to pathogenic microorganisms, strain $W5-1^T$ was isolated from Korean fermented-food Kimchi. The isolate was Gram-staining-variable, strictly aerobic, rod-shaped, endospore-forming, and motile with peritrichous flagella. It grew at $15-40^{\circ}C$, at pH 6.0-10.0, and in the presence of 0-4% NaCl. Strain $W5-1^T$ could hydrolyze esculin and xylan, and assimilate $\small{D}$-mannose, but not $\small{D}$-mannitol. Strain $W5-1^T$ showed antimicrobial activity against Listeria monocytogens, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella typhi. The G+C content of the DNA of strains $W5-1^T$ was 52.6 mol%. The predominant respiratory quinone was menaquinone-7 (MK-7) and the major cellular fatty acids were $C_{16:0}$, antieiso-$C_{15:0}$, $C_{18:0}$, and $C_{12:0}$. The strain contained meso-diaminopimelic acid in cell-wall peptidoglycan. On the basis of 16S rRNA gene sequence and phylogenetic analysis, the strain W5-1 was shown to belong to the family Paenibacillaceae and was most closely related to Paenibacillus pinihumi $S23^T$ (98.4% similarity) and Paenibacillus tarimensis $SA-7-6^T$ (96.4%). The DNA-DNA relatedness between the isolate and Paenibacillus pinihumi $S23^T$ was 8.5%, indicating that strain $W5-1^T$ represented a species in the genus Paenibacillus. On the basis of the evidence from this polyphasic study, it is proposed that strain $W5-1^T$ is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus kimchicus sp. nov. is proposed. The type strain is $W5-1^T$ (=KACC $15046^T$ = $LMG 25970^T$).

Identification of a new marine bacterium Ruegeria sp. 50C-3 isolated from seawater of Uljin in Korea and production of thermostable enzymes (대한민국 울진 연안 해양에서 분리한 해양 미생물 Ruegeria sp. 50C-3의 동정 및 내열성 효소 생산)

  • Chi, Won-Jae;Kim, Jong-Hee;Park, Jae-Seon;Hong, Soon-Kwang
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.344-351
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    • 2016
  • A marine bacterium, designated as strain 50C-3, was isolated from a seawater sample collected from the East Sea of South Korea. The strain is a Gram-negative, aerobic, yellow colored polar-flagellated bacterium that grows at $20-50^{\circ}C$ and pH 5.5-8.5. Optimal growth occurred at $40-50^{\circ}C$, at pH 6.5-7.5, and in the presence of 2% (w/v) NaCl. Based on 16S rRNA gene sequence similarity, the isolate was considered to represent a member of the genus Ruegeria. The result of this analysis showed that strain 50C-3 shared 99.4% and 96.98% sequence similarity with Ruegeria intermedia CC-GIMAT-$2^T$ and Ruegeria lacuscaerulensis ITI-$1157^T$, respectively. Furthermore, strain 50C-3 showed clear differences from related strains in terms of several characteristics such as motility, carbon utilization, enzyme production, etc. The DNA G+C content was 66.7 mol%. Chemotaxonomic analysis indicated ubiquinone-10 (Q-10) as the predominant respiratory quinone. Based on phenotypic, chemotaxonomic, and phylogenetic characteristics, the isolate represents a novel variant of the Ruegeria intermedia CC-GIMAT-$2^T$, for which we named Ruegeria sp. 50C-3 (KCTC23890=DSM25519). Strain 50C-3 did not produce cellulase and agarase, but produced alkaline phosphatase, ${\alpha}$-galactosidase, and ${\beta}$-galactosidase. The three enzymes showed stable activities even at $50^{\circ}C$ and thus regarded as thermostable enzymes. Especially, the ${\beta}$-galactosidase activity enhanced by 1.9 times at $50^{\circ}C$ than that at $37^{\circ}C$, which may be very useful for industrial application.

Cis-acting Replication Element Variation of the Foot-and-mouth Disease Virus is Associated with the Determination of Host Susceptibility (구제역바이러스의 숙주 특이성 결정에 연관되어있는 구제역바이러스 cis-acting replication element 변이 분석 연구)

  • Kang, Hyo Rin;Seong, Mi So;Ku, Bok Kyung;Cheong, JaeHun
    • Journal of Life Science
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    • v.30 no.11
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    • pp.947-955
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    • 2020
  • The foot-and-mouth disease virus (FMDV), a member of the Aphthovirus genus in the Picornaviridae family, affects wild and domesticated ruminants and pigs. During replication of the FMDV RNA (ribonucleic acid) genome, FMDV-encoding RNA polymerase 3D acts in a highly location-specific manner. This suggests that specific RNA structures recognized by 3D polymerase within non-coding regions of the FMDV genome assist with binding during replication. One such region is the cis-acting replication element (CRE), which functions as a template for RNA replication. The FMDV CRE adopts a stem-loop conformation with an extended duplex stem, supporting a novel 15-17 nucleotide loop that derives stability from base-stacking interactions, with the exact RNA nucleotide sequence of the CRE producing different RNA secondary structures. Here, we show that CRE sequences of FMDVs isolated in Korea from 2010 to 2017 exhibit A and O genotypes. Interestingly, variations in the RNA secondary structure of the Korean FMDVs are consistent with the phylogenetic relationships between these viruses and reveal the specificity of FMDV infections for particular host species. Therefore, we conclude that each genetic clade of Korean FMDV is characterized by a unique functional CRE and that the evolutionary success of new genetic lineages may be associated with the invention of a novel CRE motif. Therefore, we propose that the specific RNA structure of a CRE is an additional criterion for FMDV classification dependent on the host species. These findings will help correctly analyze CRE sequences and indicate the specificity of host species for future FMDV epidemics.

Molecular Cloning and Sequence Analysis of Coelomic Cytolytic Factor-like Gene from the Midgut of the Earthworm, Eisenia Andrei (줄지렁이 중장에서 분리한 Coelomic cytolytic factor-유사 유전자의 클로닝 및 염기서열 분석에 관한 연구)

  • Baek, Nam Sook;Lee, Myung-Sik;Park, Sang-Kil;Kim, Dae-hwan;Tak, Eun-Sik;Ahn, Chi-Hyun;Sun, Zhenjun;Park, Soon Cheol
    • Journal of the Korea Organic Resources Recycling Association
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    • v.16 no.4
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    • pp.64-73
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    • 2008
  • The cDNA of CCF (coelomic cytolytic factor)-like gene (EC 3.2.1.16), a kind of glycosyl hydorlase, was isolated and cloned from the midgut of the earthworm Eisenia anderi. The size of nucleotide sequence appeared to be 1,152 bp and its predicted coding region was composed of 384 amino acid residues including the initiation methionine. The 17 residues at N-terminal end in the deduced amino acid sequence were regarded to be a signal peptide. Based on the amino acid sequence analysis, it appeared that this CCF-like protein could belong to glycosyl hydrolase family 16 (GHF16) and showed a high sequence homology of about 79~99% with CCF and CCF-like proteins from other earthworm species. The CCFs and CCF-like proteins from various earthworm species exhibited a 100% homology in the polysacchride-binding motif and glucanase motif. It has been reported that the CCFs isolated from E. fedita appeared to show a broader pattern recognition specificity than those from other earthworm species because this species resides in decaying organic matter showing very high microbial activity, implying that CCF-like protein isolated in this study from E. andrei might exhibit a broad substrate specificity that is a useful characteristic for industrial application. A phylogenetic analysis using the deduced amino acid sequences of CCF-related proteins through the BLASTX revealed that GHF16 families could be divided into three groups of metazoa, viriplantae and eubacteria subfamily. Subsequently the CCF-related proteins of metazoa subfamily could clearly be subgroup into lophotrochozoan and edysozoan type including a deuterostome origin. Further understanding of the biological properties of E. andrei CCF-like protein should be addressed to regulate the ${\beta}$-D-glucan hydrolysis and production for the industrial uses.

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Biochemical Characterization of a Novel Thermostable Esterase from the Metagenome of Dokdo Islets Marine Sediment (독도 심해토 메타게놈 유래 신규 내열성 에스테라아제의 생화학적 특성규명)

  • Lee, Chang-Muk;Seo, Sohyeon;Kim, Su-Yeon;Song, Jaeeun;Sim, Joon-Soo;Hahn, Bum-Soo;Kim, Dong-Hern;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.63-70
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    • 2017
  • A functional screen of 60,672 fosmid metagenomic clones amplified from marine sediment obtained from the Dokdo islets in Korea identified the gene EstES1, whose product, EstES1, displayed lipolytic properties on tributyrin-supplemented media. EstES1 is a 576 amino acid protein with a predicted molecular weight of 59.4 kDa including 37 N-terminal leader amino acids. EstES1 exhibited the highest sequence similarity (44%) to a carboxylesterase found in Haliangium ochraceum DSM14365. Phylogenetic analysis indicated that EstES1 belongs to a currently uncharacterized family of lipases. Within the conserved domain, EstES1 retains the catalytic triad that consists of the consensus penta-peptide motif, GESAG. EstES1 demonstrated a broad substrate specificity toward the long acyl group of ethyl esters (C2-C12), and its optimal activity was recorded toward p-Nitrophenyl butyrate (C4) at pH 9.0 and $40^{\circ}C$ (specific activity of 255.4 U/mg). The enzyme remained stable in the ranges of $60-65^{\circ}C$ and pH 9.0-10.5 and in the presence of methanol, ethanol, isopropanol, and dimethyl sulfoxide. Therefore, EstES1 has potential for use in industrial applications involving high temperature, organic solvents, and/or alkaline conditions.

Sex Pheromone Trapping of Spodoptera frugiperda (Noctuidae: Lepidoptera) in Korea and the Distribution of Intraspecies-specific Single Nucleotide Polymorphisms in the Cytochrome c Oxidase Subunit 1 (CO1) (성페로몬을 이용한 열대거세미나방 포획과 시토크롬 옥시다제 1(CO1)에서 종내 변이군 특이적 단일염기다형성 분포)

  • Seo, Bo Yoon;Jung, Jin Kyo;Lee, Gwan-Seok;Yang, Chang Yeol;Cho, Jumrae;Kim, Yang Pyo
    • Korean journal of applied entomology
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    • v.59 no.3
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    • pp.217-231
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    • 2020
  • In 2019, the sex pheromones of Spodoptera frugiperda were used to examine moth trapping in cornfields in Gochang, Korea. Four types of traps were prepared, two funnel-types and two delta-types, each baited with 300 or 1000 ㎍ of a two-component (2C) blend of synthetic sex pheromones [100% (Z)-9-tetradecenyl acetate (Z9-14Ac) and 2% (Z)-7-dodecenyl acetate (Z7-12Ac)]. The greatest number of S. frugiperda were captured in the 300 ㎍ funnel-type trap (first catch: August 6). Large numbers of Mythimna loreyi (a non-target) were also caught in the funnel-type traps. Two wing-type traps were baited with 1000 ㎍ of the 2C blend or a four-component (4C) blend [100% (Z)-9-tetradecenyl acetate, 8% (Z)-11-hexadecenyl acetate (Z11-16Ac), 2% (Z)-7-dodecenyl acetate, and 1% (Z)-9-dodecenyl acetate (Z9-12Ac)] and the capture efficiency was assessed. Low numbers of S. frugiperda were captured regardless of the blend, and more M. loreyi were captured using the 4C blend. The two intraspecies groups clustered separately in a phylogenetic tree constructed using partial sequences (1004 bp) of cytochrome c oxidase subunit 1 (CO1). Of the 70 S. frugiperda captured in the pheromone traps, 66 belonged to CO1-RS (CO1 rice-strain) and 4 to CO1-CS (CO1 corn-strain). Twelve consistent single nucleotide polymorphisms (SNPs) were identified in CO1 between the CO1-RS and CO1-CS groups of S. frugiperda. Of the 73 S. frugiperda, 4 had the same SNP pattern as the CO1-CS group (including the corn strain) and 69 had the same SNP pattern as the CO1-RS group (including the rice strain).

Physical Characteristics and Microsatellite Polymorphisms in Miryang Native Dogs (밀양지방 토종개의 형태학적 특징 및 유전적 다양성 연구)

  • Cho, Byung-Wook;Cho, Gil-Jae
    • Journal of Life Science
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    • v.16 no.4
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    • pp.626-631
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    • 2006
  • This study was carried out to investigate the physical characteristics (height, body lenght, chest depth, head type, ear type, body color, eye type and tail type) and genetic diversity using 15 microsatellite DNA markers (PEZ 1, 5, 8, 10, 11, 12, 13, 15, 16, 17, 20, 21, FHC 2010, FHC 2054 and FHC 2079) in 44 random Miryang native dogs(6 months${\sim}$12 years old). The height, body lenght, and chest depth of Miryang native dogs were 43-55 cm(mean 49.5 cm), 45-60 cm(mean 54.3 cm), and 50-64 cm(mean 57.9 cm), respectively. Miryang native dog was medium sized. The head and eye type were reverse-triangle(100%), triangle (90.9%) and newborn moon(9.1%), respectively. Most of body color had white coat color(93.2%), light pink tongue color(100%), light black anal color(90,9%) and pink claw color(100%). The ear type showed erect ear(100%), and half-curled(56.8%), upward(34.1%), curled(9.1%) in tail type, respectively. Number of alleles observed at a single locus ranged from 2 (PEZ 21 and FHC 2010) to 14 (PEZ 13), with average number of alleles per locus of 6.13. The expected heterozygosities of 15 microsatellite loci were estimated based on gene frequencies. The highest expected heterozygosity, 0.863 was estimated in PEZ 13 locus and the lowest, 0.455 in PEZ 21 and FHC 2010 locus. And the mean expected heterozygosity of 15 microsatellite markers was calculated as 0.635. Polymorphic information content (PIC) values were ranged from 0.348 (PEZ 21 and FHC 2010) to 0.837 (PEZ 13), and the mean PIC value was calculated as 0.570. Of the 15 markers, PEZ 10, PEZ 13, PEZ 17 and FHC 2054 loci have relatively high PIC value (> 0.7) in Miryang native dog. In order to determine the efficieney of parentage control, exclusion probabilities (PE) were calculated for each allele. The highest PE 1 and PE 2 in PEZ 13 locus was caculated to 0.548 and 0.710, respectively. And the total exclusion power in PE 1 and PE 2 was calculated to 0.9895 and 0.9996, respectively. These results can give basic information for perservation and research in Miryang native dog, and phylogenetic relationships of the Korean native dog and Asian dog breeds.

Enhanced Production of Carboxymethylcellulase by a Newly Isolated Marine Microorganism Bacillus atrophaeus LBH-18 Using Rice Bran, a Byproduct from the Rice Processing Industry (미강을 이용한 해양미생물 Bacillus atrophaeus LBH-18 유래의 carboxymethylcellulase 생산의 최적화)

  • Kim, Yi-Joon;Cao, Wa;Lee, Yu-Jeong;Lee, Sang-Un;Jeong, Jeong-Han;Lee, Jin-Woo
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1295-1306
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    • 2012
  • A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus atrophaeus. This species was designated as B. atrophaeus LBH-18 based on its evolutionary distance and the phylogenetic tree resulting from 16S rDNA sequencing and the neighbor-joining method. The optimal conditions for rice bran (68.1 g/l), peptone (9.1 g/l), and initial pH (7.0) of the medium for cell growth was determined by Design Expert Software based on the response surface method; conditions for production of CMCase were 55.2 g/l, 6.6 g/l, and 7.1, respectively. The optimal temperature for cell growth and the production of CMCase by B. atrophaeus LBH-18 was $30^{\circ}C$. The optimal conditions of agitation speed and aeration rate for cell growth in a 7-l bioreactor were 324 rpm and 0.9 vvm, respectively, whereas those for production of CMCase were 343 rpm and 0.6 vvm, respectively. The optimal inner pressure for cell growth and production of CMCase in a 100-l bioreactor was 0.06 MPa. Maximal production of CMCase under optimal conditions in a 100-l bioreactor was 127.5 U/ml, which was 1.32 times higher than that without an inner pressure. In this study, rice bran was developed as a carbon source for industrial scale production of CMCase by B. atrophaeus LBH-18. Reduced time for the production of CMCase from 7 to 10 days to 3 days by using a bacterial strain with submerged fermentation also resulted in increased productivity of CMCase and a decrease in its production cost.