• Title/Summary/Keyword: r-DNA

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Phylogenetic Analysis of Caterpillar Fungi by Comparing ITS 1-5.8S-ITS 2 Ribosomal DNA Sequences

  • Park, Joung-Eon;Kim, Gi-Young;Park, Hyung-Sik;Nam, Byung-Hyouk;An, Won-Gun;Cha, Jae-Ho;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.29 no.3
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    • pp.121-131
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    • 2001
  • This study was carried out to identify the phylogenetic relationships among several caterpillar fungi by comparing the sequences of internal transcribed spacer regions(ITS1 and ITS2) and 5.8S ribosomal DNA(rDNA) repeat unit. The sequences of ITS1, ITS2, and the 5.8S rDNA from 10 strains of Cordyceps species, 12 strains of Paecilomyces, 3 strains of Beauveria, 2 strains of Metarhizium and 1 strains of Hirsutella were amplified, determined and compared with the previously known Cordyceps species. The sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites could be found. In the phylogenetic tree, the species generally divided into three clusters, supported by their morphology and/or host ranges. The 5.8S rDNA and TTS1 sequences among 10 species of Cordyceps militaris were identical and only one base pair in ITS2 sequence was different. Cordyceps sinensis and Cordyceps ophioglossoides were also clearly different, although they belonged to the same cluster. The Geniank database search of species revealed sister taxa of an entomogenous fungus. Metarhizium was used as an putgroup in all taxa.

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Effects of parental folate deficiency on the folate content, global DNA methylation, and expressions of FR${\alpha}$, IGF-2 and IGF-1R in the postnatal rat liver

  • Mejos, Karen Kay;Kim, Hye Won;Lim, Eun Mi;Chang, Namsoo
    • Nutrition Research and Practice
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    • v.7 no.4
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    • pp.281-286
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    • 2013
  • We examined the effect of parental folate deficiency on the folate content, global DNA methylation, folate receptor-alpha (FR${\alpha}$), insulin-like-growth factor-2 (IGF-2) and -1 receptor (IGF-1R) in the liver and plasma homocysteine in the postnatal rat. Male and female rats were randomly fed a folic acid-deficient (paternal folate-deficient, PD and maternal folate-deficient, MD), or folic acid-supplemented diet (paternal folate-supplemented, PS and maternal-folate-supplemented, MS) for four weeks. They were mated and grouped accordingly: $PS{\times}MS$, $PS{\times}MD$, $PD{\times}MS$, and $PD{\times}MD$. Pups were killed on day 21 of lactation. The hepatic folate content was markedly reduced in the $PD{\times}MD$ and $PS{\times}MD$ and $PD{\times}MS$ as compared with the $PS{\times}MS$ group. The hepatic global DNA methylation was decreased in the $PD{\times}MS$ and $PS{\times}MD$ groups as much as in the $PD{\times}MD$ group, and all the three groups were significantly lower as compared to the $PS{\times}MS$ group. There were no significant differences in the hepatic FR${\alpha}$, IGF-2 and IGF-1R expressions among the groups. Positive correlations were found between the hepatic folate content and global DNA methylation and protein expressions of FR${\alpha}$, IGF-2 and IGF-1R, whereas an inverse correlation was found between hepatic folate content and plasma homocysteine level in the 3-week-old rat pup. The results of this study show that both paternal and maternal folate deficiency at mating can influence the folate content and global DNA methylation in the postnatal rat liver.

Application of the Alkaline Comet Assay for Detecting Oxidative DNA Damage in Human Biomonitoring (인체 산화적 DNA손상에 대한 Human Biomonitoring도구로서 Alkaline Comet Assay의 활용 가능성 연구)

  • 박은주;강명희
    • Journal of Nutrition and Health
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    • v.35 no.2
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    • pp.213-222
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    • 2002
  • The alkaline comet assay has been used with increasing popularity to investigate the level of DNA damage in biomonitoring studies within the last decade in Western countries. The purpose of this study was to evaluate the usefulness of the alkaline comet assay as a biomarker of oxidative DNA damage for monitoring in the Korean population, and also to evaluate the effect of nutritional status and lifestyle factors on H2O2 induced oxidative DNA damage measured by the alkaline comet assay in human lymphocytes. The study population consisted of 61 healthy Korean male volunteers, aged 20-28. Epidemiological background data including dietary habits, smoking habits and anthropometrical measurements were collected through personal interviews. After blood collection, the comet assay in peripheral lymphocytes and plasma lipids analysis was carried out and the results analyzed. Tail moment (TM) and tail length (TL) of the comet assay were use\ulcorner to measure DNA damage in the lymphocytes of the subjects. Statistically significant (p < 0.05) positive correlations were observed between DNA damage (TM or TL) and smoking habits expressed as cigarettes smoked per day and pack years (r = 0.311 and 0.382 for TM, r = 0.294 and 0.350 for TL, respectively). There were also significant positive correlations between DNA damage parameter and waist-hip ratio. Higher plasma triglyceride levels were associated with increased damage to DNA. There were no correlations between the consumption frequencies of vegetables and DNA damage to the subjects. However, consumption frequencies of fruit and fruit juice intake were inversely associated with the TM and TL. The results indicate that die comet assay is a simple, rapid and sensitive method for detecting lymphocyte DNA damage induced by cigarette smoking. Consumption of fruit or fruit juices could potentiall modify the damaged DNA in the human peripheral lymphocytes of young Korean men.

Phylogenetic Diversity of Bacterial Community Inhabited in Callyspongia elegans (해면 Callyspongia elegans에 서식하는 세균군집의 계통학적 다양성)

  • Park, So-Hyun;Kim, Ji-Young;Kim, Young-Ju;Heo, Moon-Soo
    • Korean Journal of Microbiology
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    • v.50 no.2
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    • pp.152-157
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    • 2014
  • The aim of this study was to investigate the bacterial community inhabited in Callyspongia elegans. Marine bacteria were isolated from the marine sponge C. elegans using marine agar. The resulting 112 isolated pure cultures were then used for further study. They were characterized by determining morphological characteristics through Gram's staining and morphological observation. The colony pigments of bacterial isolates were characterized as yellow, brown, ivory, and white. Thirty-seven strains were found to be Gram-positive and 75 strains were Gram-negative. Seventy-nine strains were coccus-shaped, while 16 strains were rod-shaped. On the basis of the results of the comparative analyses of 16S rDNA gene sequences, the 112 isolated bacteria were divided into 5 major groups: Alphaproteobacteria (39%), Gammaproteobacteria (22%), Actinobacteria (14%), Fimicutes (9%), and Bacteroidetes (6%). It is strongly suggested that fifteen isolates are candidates for a new genera or species, based on the analyses of 16S rDNA gene sequences.

Identification and Characterization of the Causal Organism of Gummy Stem Blight in the Muskmelon (Cucumis melo L.)

  • Choi, In-Young;Choi, Jang-Nam;Choi, Dong-Chil;Sharma, Praveen Kumar;Lee, Wang-Hyu
    • Mycobiology
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    • v.38 no.3
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    • pp.166-170
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    • 2010
  • Gummy stem blight is a major foliar disease of muskmelon (Cucumis melo L.). In this study, morphological characteristics and rDNA internal transcribed spacer (ITS) sequences were analyzed to identify the causal organism of this disease. Morphological examination of the Jeonbuk isolate revealed that the percentage of monoseptal conidia ranged from 0% to 10%, and the average length $\times$ width of the conidia was 70 ($\pm$ 0.96) $\times$ 32.0 ($\pm$ 0.15) ${\mu}m$ on potato dextrose agar. The BLAST analysis showed nucleotide gaps of 1/494, 2/492, and 1/478 with identities of 485/492 (98%), 492/494 (99%), 491/494 (99%), and 476/478 (99%). The similarity in sequence identity between the rDNA ITS region of the Jeonbuk isolate and other Didymella bryoniae from BLAST searches of GenBank was 100% and was 95.0% within the group. Nucleotide sequences of the rDNA ITS region from pure culture ranged from 98.2% to 99.8%. Phylogenetic analysis with related species of D. bryoniae revealed that D. bryoniae is a monophyletic group distinguishable from other Didymella spp., including Ascochyta pinodes, Mycosphaerella pinodes, M. zeae-maydis, D. pinodes, D. applanata, D. exigua, D. rabiei, D. lentis, D. fabae, and D. vitalbina. Phylogenetic analysis, based on rDNA ITS sequence, clearly distinguished D. bryoniae and Didymella spp. from the 10 other species studied. This study identified the Jeonbuk isolate to be D. bryoniae.

Estimation of Distribution of a Commensal Thermophile in Soil by Competitive Quantitative PCR and Terminal Restriction Fragment Length Polymorphism Analysis

  • Rhee, Sung-Keun;Hong, Seung-Pyo;Bae, Jin-Woo;Jeon, Che-Ok;Lee, Seung-Goo;Song, Jae-Jun;Poo, Ha-Ryoung;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.940-945
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    • 2001
  • Symbiobacterium toebii has been previously reported as a novel commensal thermophile exhibiting a commensal interaction with thermophilic Geobacillus sp. SK-1. We investigated the distribution of this commensal thermophile in various soils using molecular methods, such as quantitative PCR and terminal restriction fragment polymorphism analysis. Based on a nested competitive quantitative PCR the 16S rDNA of the commensal thermophile was only detected in compost soils at about $1.0{\times}10^4$ cpoies per gram of soil, corresponding to $0.25{\times}10^4$ cells per gram of soil. However, in an enrichment experiment at $60^{\circ}C$, about $1.0{\times}10^8$ copies of 16S rDNA molecules were detected per ml of enriched culture broth for all the soils, and more than 0.1 mM indole accumulated as the product of commensal bacterial growth. When incubated at $30^{\circ}C$, neither the 16S rDNA of the commensal bacterium nor any indole accumulation was detected. Accordingly, even though the 16S rDNA of the bacterium was only detected in the compost soils by a nested PCR, the presence of the 16S rDNA molecules of commensal thermophile and accumulation of indole in all the enriched cultures appeared to indicate that the commensal thermophile is widely distributed in various soils.

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Comparison Between Phylogenetic Relationships Based on 18S rDNA Sequences and Growth by Salinity of Chlorella-like Species (Chlorophyta)

  • Lee, Hye-Jung;Hur, Sung-Bum
    • Fisheries and Aquatic Sciences
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    • v.15 no.2
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    • pp.125-135
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    • 2012
  • This study was carried out to understand the correlation between phylogenetic relationships based on 18S rDNA sequences and growth by salinity of Chlorella-like species. The 18S rDNA sequences of 71 Chlorella-like species which were mainly collected from Korean waters were analyzed. The 18S rDNA sequences of Chlorella-like species were divided into three groups (group A, B and C) and group B was further divided into three subgroups (subgroup B-1, B-2 and B-3). Thirty-seven Chlorella-like species in group A grew well at high salinity (32 psu) but the other groups grew well in freshwater. The sequence identities of the species in group A and B were 97.2-99.5%, but those of 6 species in group C ("Chlorella" saccharophila), which contained group I intron sequences region were 75.0-75.4%. Two representative species of each group were cultured at different salinities (0, 16 and 32 psu) to examine the correlation between the molecular phylogenetic groups and the phenotypic characteristics on cell growth and size by different salinities. The size of cell cultured at different salinities varied according to the species of each molecular phylogenetic group. The size of "Chlorella" saccharophila in group C was bigger and more obviously elliptical rather than that of the other Chlorella-like species. Considering the results on molecular and phenotypic characteristics, the group A and B belonged to Chlorellaceae, but group C was distinctly different from them.

The Diversity of Heterotrophic Bacteria Isolated from Intestine of Starfish(Asterias amurensis) by Analysis of 16S rDNA Sequence (16S rDNA염기서열에 의한 불가사리(Asterias amurensis) 장내에서 분리된 종속영양세균 군집의 다양성)

  • Choi, Gang-Guk;Lee, Oh-Hyung;Lee, Geon-Hyoung
    • The Korean Journal of Ecology
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    • v.26 no.6
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    • pp.307-312
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    • 2003
  • To study the diversity of heterotrophic bacteria isolated from intestine of starfish, Asterias amurensis, we collected starfishes from the coastal area near Jangheung-Gun, Jeollanam-Do, Korea during July, 2000. Population density and bacterial diversity in the intestine of starfish were measured. The results were as follows; The population densities of heterotrophic bacteria in the intestine of starfish were 8.65${\pm}$0.65${\times}10^3\;dfu\;g^{-1}$. Gram positive bacteria occupied 59% among 29 isolates. The community structure of dominant heterotrophic bacteria in the intestine of starfish consisted of Bacillaceae in the low G+C gram positive bacteria subphylum, Microbacteriaceae in the high G+C gram positive bacteria subphylum, and Alteromonadaceae in ${\gamma}$-Proteobacteria subphylum. Among eight strains of Bacillus spp., three strains showed more than 97% identity, but five strains showed about 90% identity with type strain on the basis of partial 16S rDNA sequence.

Characterization of a Ligninase Producing Strain, Serratia marcescens HY-5 isolated from Sympetrum dopressiusculum (고추좀잠자리 (Sympetrum depressiusculum)로부터 분리한 리그닌 분해균주, Serratia marcescens HY-5의 특성)

  • Kim, Ki-Duck;Park, Doo-Sang;Shin, Dong-Ha;Han, Bo-Na;Oh, Hyun-Woo;Youn, Young-Nam;Park, Ho-Yong
    • Korean journal of applied entomology
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    • v.45 no.3 s.144
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    • pp.301-307
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    • 2006
  • A ligniolytic bacterial strain was isolated from the digestive tract of a red dragonfly, Sympetrum dopressiusculum. It was identified as a Serratia marcescens HY-5 by 16S rDNA sequence analysis and physiological and biochemical analysis. The isolated strain showed proportional increase of ligninolytic activity to the cell growth in the culture media which include lignin compounds. It showed about 25-45% decomposition of lignin compound by 48 hr incubation especially, showed effective decomposition of monomer lignin compounds, vanillin and guaiacol, and a dimer, dealkaline lignin. PCR amplification of 16S rDNA followed by denaturing gradient gel electrophoresis analysis showed high density of S. marcescens HY-5 in the gut of the S. depressiusculum at both gut samples which collected at different site.

Effect of Farnesyltransferase Inhibitor R115777 on Mitochondria of Plasmodium falciparum

  • Ha, Young Ran;Hwang, Bae-Geun;Hong, Yeonchul;Yang, Hye-Won;Lee, Sang Joon
    • Parasites, Hosts and Diseases
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    • v.53 no.4
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    • pp.421-430
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    • 2015
  • The parasite Plasmodium falciparum causes severe malaria and is the most dangerous to humans. However, it exhibits resistance to their drugs. Farnesyltransferase has been identified in pathogenic protozoa of the genera Plasmodium and the target of farnesyltransferase includes Ras family. Therefore, the inhibition of farnesyltransferase has been suggested as a new strategy for the treatment of malaria. However, the exact functional mechanism of this agent is still unknown. In addition, the effect of farnesyltransferase inhibitor (FTIs) on mitochondrial level of malaria parasites is not fully understood. In this study, therefore, the effect of a FTI R115777 on the function of mitochondria of P. falciparum was investigated experimentally. As a result, FTI R115777 was found to suppress the infection rate of malaria parasites under in vitro condition. It also reduces the copy number of mtDNA-encoded cytochrome c oxidase III. In addition, the mitochondrial membrane potential (${\Delta}{\Psi}m$) and the green fluorescence intensity of MitoTracker were decreased by FTI R115777. Chloroquine and atovaquone were measured by the mtDNA copy number as mitochondrial non-specific or specific inhibitor, respectively. Chloroquine did not affect the copy number of mtDNA-encoded cytochrome c oxidase III, while atovaquone induced to change the mtDNA copy number. These results suggest that FTI R115777 has strong influence on the mitochondrial function of P. falciparum. It may have therapeutic potential for malaria by targeting the mitochondria of parasites.