• Title/Summary/Keyword: 16s rRNA 유전자

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Characteristics and Antibiotic Susceptibility of Imipenem-Resistant Clinical Isolates Producing Carbapenemase (Carbapenemase를 생산하는 imipenem 내성 세균의 특성 및 항생제 감수성)

  • Choe, Han-Na;Park, Chul;Kim, Hyung-Rak;Baik, Keun-Sik;Kim, Se-Na;Seong, Chi-Nam
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1214-1220
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    • 2010
  • Imipenem-resistant bacteria were isolated from clinical specimens taken from hospitalized patients in Suncheon, Korea. Fifty-four isolates were phylogenetically analyzed based on 16S rRNA gene and gyrB gene sequence comparisons. Isolates were affiliated with Pseudomonas aeruginosa (30 strains; 55.6%), Acinetobacter baumannii (21; 38.9%), Enterobacter hormaechei (2) and Pseudomonas putida (2). Twenty-two isolates produced metallo-$\beta$-lactamase (MBL); 12 Acinetobacter baumannii strains, 7 Pseudomonas aeruginosa strains, 2 P. putida strains and 1 Enterobacter hormaechei strain. Antibiotic susceptibility of the isolates was determined using the disc diffusion method and Vitek system. Strains producing metallo-$\beta$-lactamase (type IMP & VIM) were more resistant to antibiotics ceftazidime, aztreonam, amikacin and gentamicin than to strains producing OXA and SHV type of $\beta$-lactamase.

Effect of Obesity and Diabetes on Alzheimer's APP Gene Expression in Mouse Adipose Tissues (비만 및 당뇨가 생쥐 지방조직에서의 Alzheimer's APP 유전자 발현에 미치는 영향)

  • Kim, Jin-Woo;Lee, Yong-Ho
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1012-1018
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    • 2010
  • The aim of this study was to determine whether Alzheimer's amyloid precursor protein (APP) is dysregulated in adipose tissues of C57BL/6 male mice by high-fat diet (HFD) induced obesity, aging, or streptozotocin (STZ)-induced diabetes. APP mRNA expression was examined by quantitative real-time PCR (QPCR) in subcutaneous (SAT) and epididymal adipose tissues (EAT) from mice in 8 different condition groups. By combining conditions of age (16 weeks/26 weeks of age), diet (normal diet (ND)/high-fat diet), and induction of diabetes (non-diabetic/diabetic), 88 mice were divided into 8 different groups. QPCR demonstrated that APP expression in SAT was significantly increased by about two-fold in HFD-induced obese mice compared to both 16 week-old and 26 week-old mice in the ND group (16 weeks p=0.001; 26 weeks p<0.0001), but no changes in EAT was found. Particular effects of aging on APP gene expression were not observed in either adipose tissue depots. Significantly decreased APP expression was found in SAT in STZ-induced diabetic mice fed on ND or HFD at 16 weeks of age (ND p<0.05; HFD p<0.01). Linear regression analysis demonstrated that APP expression levels correlated with body weight in both the non-diabetic group (R=0.657, p<0.0001, n=39) and the diabetic group (R=0.508, p=<0.0001, n=49), but did not correlate with plasma glucose levels, which suggests that decreased APP expression in STZ-induced diabetic mice is most likely due to weight loss rather than hyperglycemia. These data confirm APP dysregulation by weight changes in humans and suggest a possible role linking midlife obesity with the later development of amyloidogenesis in the brain of older patients with Alzheimer's disease.

Development of Recombinant Escherichia coli Expressing Rhodotorula glutinis Epoxide Hydrolase (Rhodotorula glutinis의 epoxide hydrolase 고효율 발현 유전자 재조합 Escherichia coli 생촉매 개발)

  • Lee Soo-Jung;Kim Hee-Sook
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.415-419
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    • 2006
  • The epoxide hydrolase (EH) of Rhodotorula glutinis which has a high enantioselectivity against aromatic epoxide substrates was expressed to high levels in Escherichia coli based on codon usage. We analysed the Preference of codon usage between the yeast, R. glutinis, and bacteria, E. coli. E. coli, Rosetta(DE3)pLysS, harbors pRARE plasmid with tRNA genes for rare-codons was employed as a host strain. The recombinant E. coli expressing R. glutinis EH showed an enhanced enantioselective hydrolysis activity toward racemic styrene oxide. Enantiopure (S)-styrene oxide with a high enantiopurity of 99% ee (enantiomeric excess) was obtained from racemic substrates.

Cloning and Characterization of Mannanase Gene from Bacillus subtilis WL-8 (Bacillus subtilis WL-8의 Mannanase 유전자 클로닝과 특성분석)

  • Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.207-212
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    • 2010
  • A bacterium producing the extracellular mannanase was isolated from Korean soybean paste. The isolate WL-8 has been identified as Bacillus subtilis on the basis on its 16S rRNA sequence, morphology and biochemical properties. The mannanase productivity of strain WL-8 was increased in LB broth by addition of wheat bran. The maximum mannanase productivity was reached to approximately 20 U/ml in LB medium supplemented with 6% wheat bran. A gene encoding the mannanase of WL-8 was cloned into Escherichia coli and its nucleotide sequence was subsequently determined. The mannanase gene consisted of 1,086 nucleotides encoding a polypeptide of 362 amino acid residues. The deduced amino acid sequence was highly homologous with those of several mannanases from B. subtilis belonging to GH family 26. Reaction temperature and pH profiles were investigated using the culture filtrate and cell-free extract of the recombinant E. coli carrying a WL-8 mannanase gene, respectively. Optimal conditions for the two fractions occurred at pH 5.5 and $60^{\circ}C$. The cell-free extract showed higher mannanase activity than the culture filtrate at above $60^{\circ}C$.

Complete Mitochondrial Genome Sequence and Genetic Diversity of Duroc Breed (돼지 Duroc 품종에서 미토콘드리아 유전체 서열의 특성과 집단의 유전적 다양성)

  • Cho, 1.C.;Han, S.H.;Choi, Y.L.;Ko, M.S.;Lee, J.G;Lee, J.H;Jeon, J .T
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.937-946
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    • 2004
  • Duroc is widely used to improve the meat quality and productivity. To elucidate the phylogenetic relation and the sequence specificity for the maternal property, the complete sequence of mitochondrial genome was determined and the population diversity of Duroc was investigated in this study. The length of mtDNA tested is 16,584-bp. There are several insertion/deletion mutations in the control region and coding regions for tRNA and rRNA, respectively, but not in peptide-coding regions. Four peptide-coding genes(COⅡ, COⅢ, ND3 and ND4) showed incomplete termination codon sequences such as T--, and two(ND2 and ND4L) did alternative initiation codons(AIC), respectively. Especially, the initiation codon sequences of ND2 gene were polymorphic in this population. Polymorphisms were detected in 11-bp duplication motif within control region as well as ND2 and CYTB. Variation patterns observed from the tests on three mtDNA regions were linked completely and then two haplotypes obtained from combining the data dividing this population. Duroc mtDNA is observed at the European pig cluster in the phylogenetic tree, however, the results from the population analyses supported previous opinions. This study suggests that the breed Duroc was mainly originated from the European pig lineage, and Asian lineage was also used to form the pig breed Duroc as maternal progenitors.

Detection of Gram-negative Bacteria in Broad-range PCR Amplifying 16S rRNA Gene with Semi-nested Primers and Its Application in Market Milk (16S rRNA 유전자의 Semi-nested Primer를 이용한 Broad-range PCR에 의한 그람음성세균의 검출과 시유에서의 응용)

  • Choi, Suk-Ho;Choi, J.J.;Lee, S.B.
    • Journal of Animal Science and Technology
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    • v.47 no.3
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    • pp.465-474
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    • 2005
  • A two-step broad-range PCR method detecting gram-negative bacteria at the level as low as 2 CFU was developed by using primers of GNFI and GNRI and then semi-nested primer of GNF2 and GNRI. The nucleotide sequences of the primers were determined based on l6S rRNA gene. The DNA fragments of 1173 bp and 169 bp were amplified in one-step PCRs with primer sets of GNFI-GNRI and GNF2-GNRl, respectively, using template DNA from seven strains of gram-negative bacteria including Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas spp., and Acinetobacter baumaii but not from Achromobacter lyticus, Alca/igens faecalis, and five strains of gram-positive bacteria. DNA fragments of 180 bp were amplified from LTLT-pasteurized milk and UHf-pasteurized milk in the two-step PCR. The DNA fragments were amplified from LTLT-pasteurized milk which was added with Pseudomonas j/uorescens and subsequently heated at 65 $^{\circ}C$, 80 $^{\circ}C$, and 100 $^{\circ}C$ for 30 min but they were not amplified from the milk autoclaved at 121$^{\circ}C$ for 15 min. It was suggested in PCR that Pseudomonas fluorescens heated at 65 $^{\circ}C$ for 30 min in milk was more sensitive to DNase treatment than viable bacteria.

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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Antibiotic and Insecticidal Activities of Metabolites Derived From an Entomopathogenic Bacterium, Xenorhabdus nematophila, Against The Bean Bug, Riptortus clavatus (곤충병원세균(Xenorhabdus nematophila) 유래물질의 톱다리개미허리노린재(Riptortus clavatus) 장내세균 증식억제 및 살충효과)

  • Seo, Sam-Yeol;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.49 no.3
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    • pp.251-259
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    • 2010
  • A bacterial colony was isolated from the gut of the bean bug, Riptortus clavatus. From morphological and biochemical tests, the bacterial isolate showed the highest similarity to Staphylococcus succinus. DNA sequence of 16S rRNA gene of the bacterium supported the identification. Oral administration of penicillin G to adults of R. clavatus gave a dose-dependent mortality of adults of R. clavatus to adults along with significant decrease of the bacterial population in the gut. Similarly, three metabolites (benzylideneacetone, proline-tyrosine, and acetylated phenylalanine-glycine-valine) derived from an entomopathogenic bacterium, Xenorhabdus nematophila, also inhibited growth of the gut bacterial population and gave significant mortalities to R. clavatus. These results suggest that a gut bacterial population classified as Staphylococcus sp. is required for survival of R. clavatus and that the three bacterial metabolites had toxic effects on the bugs due to their antibacterial properties.

Identification and Biochemical Characterization of a New Xylan-degrading Streptomyces atrovirens Subspecies WJ-2 Isolated from Soil of Jeju Island in Korea (제주도 토양으로부터 자일란 분해 Streptomyces atrovirens subspecies WJ-2 동정 및 효소의 생화학적 특성 규명)

  • Kim, Da Som;Bae, Chang Hwan;Yeo, Joo Hong;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.512-521
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    • 2016
  • A bacterial strain was isolated from a soil sample collected on Jeju Island, Korea. The strain, designated WJ-2, exhibited a high xylanase activity, whereas cellulase activity was not detected. The 16S rRNA gene sequence of WJ-2 was highly similar to type strains of the genus Streptomyces. A neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain WJ-2 is phylogenetically related to Streptomyces atrovirens. Furthermore, DNA-DNA hybridization analysis confirmed that strain WJ-2 is a novel subspecies of Streptomyces atrovirens. The genomic DNA G+C content was 73.98 mol% and the major fatty acid present was anteiso-C15:0 (36.19%). The growth and xylanase production of strain WJ-2 were significantly enhanced by using soytone and xylan as nitrogen and carbon sources, respectively. Crude enzyme preparations from the culture broth of strain WJ-2 exhibited maximal total xylanase activities at pH 7.0 and $55^{\circ}C$. Thin-layer chromatography analysis revealed that the crude enzyme degrades beechwood xylan to yield xylobiose and xylotriose as the principal hydrolyzed end products.