• Title/Summary/Keyword: 23S rDNA

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Detection of 23S rRNA Mutation Associated with Clarithromycin Resistance in Children with Helicobacter pylori Infection (소아 Helicobacter pylori 감염에서 Clarithromycin 내성과 연관된 23S rRNA의 돌연변이)

  • Ko, Jae Sung;Yang, Hye Ran;Seo, Jeong Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.7 no.2
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    • pp.137-142
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    • 2004
  • Purpose: The resistance of H. pylori to clarithromycin is one of the major causes of eradication failure. In H. pylori, clarithromycin resistance is due to point mutation in 23S rRNA. The aims of this study were to investigate the mutation of 23S rRNA and to examine the association of cagA, vacA genotype and clarithromycin resistant genes. Methods: H. pylori DNA was extracted from antral biopsy specimens from 27 children with H. pylori infection. Specific polymerase chain reaction (PCR) assays were used for cagA and vacA. Mutations associated with clarithromycin resistance were detected by using PCR restriction fragment length polymorphism (RFLP) analysis of 23S rRNA gene. Results: A2143G mutation was detected in one case and A2144G in 4, indicating 18.5% were clarithromycin resistant. Among the total of 27, cagA was present in 25 (93%), vacA s1a/m1 in 6 (22%), s1a/m2 in 3 (11%), s1c/m1 in 16 (59%), and s1c/m2 in 1 (4%). All of the 5 clarithromycin resistant strains were cagA (+), among which 2 were s1a/m1 and 2 were s1c/m1. There was no relation between genotypes and clarithromycin resistant genes. Conclusion: Detection of H. pylori resistance to clarithromycin using PCR RFLP from biopsy specimens might be useful for the selection of antibiotics. Clarithromycin resistant genes are not associated with genotypes of cagA and vacA.

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Studies on the riboxomal RNA genes of rhizobium meliloti and bradyrhizobium japonicum (Rhizobium meliloti와 bradyrhizobium japonicum의 ribosomal RNA 유전자에 관한 연구)

  • 강홍규;김달웅;하지홍
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.312-317
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    • 1988
  • The genes for ribosomal RNA in Rhizobium meliloti and Bradyrhizobium japonicum were analyzed by southern hybridization of BamHI, EcoRI, HindIII digested chromosomal DNA with purified 5' $^{32}P$-labeled 16S and 23S rRNA. The big differences in the hybridization pattern of both rhizobia were found. The comparative results were discussed in relation to the copy number and conservativity of restriction sites in the rRNA genes of both rhizobia.

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Characteristics and Diagnostic Methods of Streptococcosis Causing Disease in Aquaculture (양식 어류에 질병을 유발하는 연쇄구균증의 특성 및 진단 방법)

  • Kim, Dong-Hwi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1118-1126
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    • 2018
  • In this study, investigated the general characteristics and diagnostic methods types of streptococcosis among various fish disease pathogens that caused a lot of economic damaged to aquaculture fish based on the previous research paper. Streptococcosis infection of fish is considered a reemerging disease affecting a variety of wild and cultured fish throughout the world. Calssifiacation of Gram positive cocci based on DNA-DNA hybridization coupled with 16S sequencing has shown that at least five different species are considered of significance as fish pathogens: Lactococcus garvieae, L. piscium, Streptococcus iniae, S. agalactiae, S. paruberis, Vagococcus salmoninarum. Symptoms of infection with streptococcosis disease such as body color change, eyeball abnormality, gill discoloration, bleeding, abdominal distension, swelling of the kidney and spleen. In addition, it usually occurs from June to October when the water temperature rise a lot of fish death. Currently, 16S rRNA, 16S-23S rRNA intergenic spacer region (ISR), Random Amplified polymorphic DNA (RAPD), Ribotyion (RT), Loop-mediated isothermal amplification (LAMP) are among the methods for diagnosing streptococcosis. Among them, the LAMP method, which is high applicable to the aquaculture farm has attracted the spotlight, but due to problems such as confirmation of results. This seems to minimize the economic loss of streptococcosis which complements the problem so that it can be easily used from the diagnosis to the results confirmation.

Genetic and Phenotypic Diversity of (R/S)-Mecoprop [2-(2-Methyl-4- Chlorophenoxy)Propionic Acid]-Degrading Bacteria Isolated from Soils

  • Lim, Jong-Sung;Jung, Mee-Kum;Kim, Mi-Soon;Ahn, Jae-Hyung;Ka, Jong-Ok
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.87-93
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    • 2004
  • Twelve mecoprop-degrading bacteria were isolated from soil samples, and their genetic and phenotypic characteristics were investigated. Analysis of 16S rDNA sequences indicated that the isolates were related to members of the genus Sphingomonas. Ten different chromosomal DNA patterns were obtained by polymerase-chain-reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences from the 12 isolates. The isolates were found to be able to utilize the chiral herbicide meco-prop as a sole source of carbon and energy. While seven of the isolates were able to degrade both (R)-and (S)-mecoprop, four isolates exhibited enantioselective degradation of the (S)-type and one isolate could degrade only the (R)-enantiomer. All of the isolates were observed to possess plasmid DNAs. When certain plasmids were removed from isolates MPll, MP15, and MP23, those strains could no longer degrade mecoprop. This compelling result suggests that plasmid DNAs, in this case, conferred the ability to degrade the herbicide. The isolates MP13, MP15, and MP24 were identified as the same strain; however, they exhibited different plasmid profiles. This indicates that these isolates acquired dif-ferent mecoprop-degradative plasmids in different soils through natural gene transfer.

Identification and Characterization of Alginate Lyase Producing Pseudomonas sp. N7151-6 (Alginate Lyase 생산 균주 Pseudomonas sp. N7151-6의 분리 및 특성)

  • Lee, Jae-Hyung;Bae, Min-Ji;Kim, Yang-Choon;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.350-354
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    • 2009
  • A Gram-negative, alginate lyase-producing bacterium was isolated from the Haeundae Coast, Korea. The isolated strain N7151-6 produced alginate lyase. The optimal temperature and pH for growth were found to be $30^{\circ}C$ and pH 8.0, respectively. This strain can be grown at the NaCl concentration of 0-7% (w/v). Analysis of 16S rDNA sequence and physiological profiling indicated that the strain N7151-6 belonged to Pseudomonas sp. The enzyme alginate lyase produced by Pseudomonas sp. N7151-6 was partially purified by ultrafiltration (MWCO= 30 kDa). The optimum pH and temperature for the activity of the purified enzyme were found to be 7.0 and $30^{\circ}C$, respectively. The enzyme was stable at the pH range of 5.0-9.0 and temperature range of $23-30^{\circ}C$. The total activity of alginate lyase produced was reached about 110 unit/L.

Occurrence of Natural Hybrid between Oplegnathus fasciatus and Oplegnathus punctatus from the South Sea of Korea (한국 남해에서 출현한 돌돔 (Oplegnathus fasciatus)과 강담돔 (Oplegnathus punctatus) 사이의 자연교잡종)

  • Kwun, Hyuck-Joon;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.22 no.3
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    • pp.201-205
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    • 2010
  • One specimen of a natural hybrid of an Oplegnathus (Oplegnathus fasciatus $\times$ Oplegnathus punctatus) was found in Tongyeong, Korea in August 2008. We, herein, describe its morphological and genetic characteristics and compare them with those of O. fasciatus and O. punctatus. In morphology, the hybrid showed many distinctive black rounded blotches on body sides and four faint vertical bars, being in those features similar to O. punctatus. Although the counts and measurements of the hybrid mostly overlapped between O. fasciatus and O. punctatus, the Oplegnathus hybrid resembled O. punctatus in the ratio of pelvic-fin length in standard length: Oplegnathus hybrid (26.7%) was closer to O. punctatus (26.4%) than to O. fasciatus (17.2~23.6%). In genetics, as a result of analysis of 510 base pair sequences of mitochondrial DNA 16S rRNA, the hybrid was closer to O. fasciatus (d=0.000~0.010) than to O. punctatus (d=0.020). Our results suggest that the natural hybridization represented by the subject specimen occurred between an O. fasciatus female and an O. punctatus male.

Marine Prokaryotic Diversity of the Deep Sea Waters at the Depth of 1500 m Off the Coast of the Ulleung Island in the East Sea (Korea) (울릉도 연안 수심 1500 m에 서식하는 해양미생물군집의 분포)

  • Kim, Mi-Kyung;Khang, Yongho
    • Korean Journal of Microbiology
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    • v.48 no.4
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    • pp.328-331
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    • 2012
  • Microbial diversity in the 1500 m depth sea waters off the coast of Ulleung island of the East Sea, Korea, was investigated. Genomic DNAs were extracted directly from the marine microbes filtered through ultramembrane filters. Pyrosequencing of 16S rDNAs of these microbes resulted in 13,029 reads, of which uncultured bacteria consisted of 54.1%, alphaproteobacteria 23.4%, and gammaproteobacteria 22.3%. Other classes such as flavobacteria, actinobacteria, and epsilonproteobacteria were distributed within 0.2% of total reads. Among the cultivable bacteria, it was found that Rhodobacteraceae family of alphaproteobacteria, Alteromonadaceae, Halomonadaceae, and Piscirickettsiaceae families of gammaproteobacteria were mostly distributed in the deep-sea waters.

Site-directed Mutagenesis Analysis Elucidates the Role of 223/227 Arginine in 23S rRNA Methylation, Which Is in 'Target Adenine Binding Loop' Region of ErmSF (위치 지정 치환 변이를 이용한 ErmSF의 '타깃 Adenine Binding Loop'을 형성하는 부위에 존재하는 223/227 Arginine 잔기의 23S rRNA Methylation 활성에서의 역할 규명)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.79-86
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    • 2012
  • ErmSF is one of the Erm family proteins which catalyze S-adenosyl-$_L$-methionine dependent modification of a specific adenine residue (A2058, E. coli numbering) in bacterial 23S rRNA, thereby conferring resistance to clinically important macrolide, lincosamide and streptogramin B ($MLS_B$) antibiotics. $^{222}FXPXPXVXS^{230}$ (ErmSF numbering) sequence appears to be a consensus sequence among the Erm family. This sequence was supposed to be involved in direct interaction with the target adenine from the structural studies of Erm protein ErmC'. But in DNA methyltarnsferase M. Taq I, this interaction have been identified biochemically and from the complex structure with substrate. Arginine 223 and 227 in this sequence are not conserved among Erm proteins, but because of the basic nature of residues, it was expected to interact with RNA substrates. Two amino acid residues were replaced with Ala by site-directed mutagenesis. Two mutant proteins still maintained its activity in vivo and resistant to the antibiotic erythromycin. Compared to the wild-type ErmSF, R223A and R227A proteins retained about 50% and 88% of activity in vitro, respectively. Even though those arginine residues are not essential in the catalytic step, with their positive charge they may play an important role for RNA binding.

Genomic DNA Methylation Status and Plasma Homocysteine in Choline- and Folate-Deficient Rats (콜린과 엽산 결핍이 흰쥐의 Genomic DNA 메틸화와 혈장 호모시스테인에 미치는 영향)

  • Mun, Ju-Ae;Min, Hye-Sun
    • Journal of Nutrition and Health
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    • v.40 no.1
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    • pp.14-23
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    • 2007
  • Elevated plasma homocysteine (Hcy) is a risk factor for cognitive dysfunction and Alzheimer disease, although the mechanism is still unknown. Both folate and betaine, a choline metabolite, play essential roles in the remethylation of Hcy to methionine. Choline deficiency may be associated with low folate status and high plasma Hcy. Alterations in DNA methylation also have established critical roles for methylation in development of the nervous system. This study was undertaken to assess the effect of choline and folate deficiency on Hcy metabolism and genomic DNA methylation status of the liver and brain. Groups of adult male Sprague Dawley rats were fed on a control, choline-deficient (CD), folate-deficient (FD) or choline/folate-deficient (CFD) diets for 8 weeks. FD resulted in a significantly lower hepatic folate (23%) (p<0.001) and brain folate (69%) (p<0.05) compared to the control group. However, plasma and brain folate remained unaltered by CD and hepatic folate reduced to 85% of the control by CD (p<0.05). Plasma Hcy was significantly increased by FD $(18.34{\pm}1.62{\mu}M)$ and CFD $(19.35{\pm}3.62{\mu}M)$ compared to the control $(6.29{\pm}0.60{\mu}M)$ (p<0.001), but remained unaltered by CD. FD depressed S-adenosylmethionine (SAM) by 59% (p<0.001) and elevated S-adenosylhomocysteine (SAM) by 47% in liver compared to the control group (p<0.001). In contrast, brain SAM levels remained unaltered in CD, FD and CFD rats. Genomic DNA methylation status was reduced by FD in liver (p<0.05) Genomic DNA hypomethylation was also observed in brain by CD, FD and CFD although it was not significantly different from the control group. Genomic DNA methylation status was correlated with folate stores in liver (r=-0.397, p<0.05) and brain (r = -0.390, p<0.05), respectively. In conclusion, our data demonsoated that genomic DNA methylation and SAM level were reduced by folate deficiency in liver, but not in brain, and correlated with folate concentration in the tissue. The fact that folate deficiency had differential effects on SAM, SAH and genomic DNA methylation in liver and brain suggests that the Hcy metabolism and DNA methylation are regulated in tissue-specific ways.

Identification of a Pathogenic Bacterium, Staphylococcus gallinarum, to Bombyx mori (누에(Bombyx mori) 무름병을 야기하는 병원성 세균 Staphylococcus gallinarum의 동정)

  • 김길호;박영진;김용균
    • Korean journal of applied entomology
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    • v.41 no.4
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    • pp.279-284
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    • 2002
  • Flacherie symptom was found in the fifth instar larvae of silkworm, Bombyx mori. The bacterial pathogen was isolated from the hemolymph of the infected silkworm and identified. The isolated bacteria caused a significant flacherie pathogenicity to the fifth instar larvae of B. mori when $5{\times}10^{6}$ cfu (colony-forming unit) of the bacteria was injected into each larva. The infected larvae began to die at 6 days after injection and resulted in complete mortality at 10 days. The bacterium was identified as Staphylococcus gallinarum based on the morphological and physiological characteristics described in Bergey's manual. This identification was further supported by the characters of carbohydrate utility analyzed from a bacterial identification system ($MicroLog^{\circledR}$) and also by the molecular structure of 165-23S rDNA internal transcribed spaces. As an insecticidal action, S. gallinarum caused hemolymph septicemia by its cytotoxic effect on the hemocytes of B. mori.