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          • KCI등재

            2010년도 국내 임상에서 분리한 다제내성 녹농균의 유전자형 조사

            김민지,차민경,이도경,강주연,박재은,김영희,박일호,신혜순,하남주,Kim,,Min,Ji,Cha,,Min,Kyeong,Lee,,Do,Kyung,Kang,,Ju,Yeon,Park,,Jae,Eun,Kim,,Young,Hee,Park,,Il,Ho,Shin,,Hea,Soon,Ha,,Nam,Joo The Microbiological Society of Korea 2012 미생물학회지 Vol.48 No.4

            녹농균은 특히 면역이 저하된 환자에게서 심각한 감염을 일으키는 그람음성의 기회감염 균주이다. 또한 carbapenem 내성 metallo-${\beta}$-lactamases (MBL)를 가진 녹농균이 한국에서 증가되는 추세로 보고되고 있다. 따라서 본 실험에서는 2차 병원인 삼육 서울 병원에서 수집된 총 92종의 임상 녹농균의 다재내성 수준을 분석하였다. 항생제에 대한 감수성은 최소억제농도(MIC) 분석에 의해 결정되었고, inhibitor-potentiated disk diffusion(IPD) 분석은 MBL 검출을 위해 수행되었다. RAPD-PCR은 임상환자에서 분리한 녹농균 계통의 유전적 유형의 특징을 밝히기 위해 사용되었다. 그 결과 임상에서 분리된 녹농균의 40.2%는 ceftazidime에 내성을, 58.7%는 meropenem에 내성을, 56.5%는 gentamicin에 내성을, 46.7%는 tobramycin에 내성을, 62.0%는 ciprofloxacin에 내성을 그리고 97.8%는 chloramphenicol에 내성을 보였다. IPD 분석에 의해 29종의 다재내성 균주로 관찰 되었고, RAPD 분석에 의해 19종은 IPM-1 유전자형을, 2종은 VIM-2 유전자형을 만들었다. MBL 유전자 검출 시험을 통해 19종의 IMP-1 생성 녹농균 중에서 16종이 유사한 유전자형을 보였고, 3종은 다른 유전자형이 관찰되었다. 임상에서 분리한 IMP-1 생성 다재내성 녹농균의 비율은 꾸준히 증가하고 있다. 이번 연구는 2010년 국내 임상에서 분리한 녹농균의 항생제 다제내성 패턴과 유전자형에 대한 정보를 제공한다. Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium that causes serious infection, particularly in immunocompromised patients. Also, P. aeruginosa possessing carbapenem-resistant metallo-${\beta}$-lactamases (MBL) has been reported with increasing frequency in Korea. We therefore analyzed the level of multidrug-resistant clinical P. aeruginosa isolated from a secondary hospital in Korea in 2010. A total of 92 isolates of P. aeruginosa were collected from Sahmyook Medical Center in 2010. Susceptibility to antimicrobial agents was determined by analysis of the minimum inhibitory concentration test; the inhibitor-potentiated disk diffusion (IPD) test was performed for MBL detection. RAPD-PCR was used for genotyping to rapidly characterize P. aeruginosa strains isolated from clinical patients. The percentages of non-susceptible isolates were as follows: 40.2% to ceftazidime, 58.7% to meropenem, 56.5% to gentamicin, 46.7% to tobramycin, 62.0% to ciprofloxacin and 97.8% to chloramphenicol. The 29 multidrug-resistant strains were screened by the IPD test: of the 21 PCR-positive isolates, 19 were IPM-1 producers and 2 were VIM-2 producers. Among the 19 IMP-1-producing P. aeruginosa isolates, 16 isolates showed similar patterns, and three different banding patterns were observed. The proportion of IMP-1-producing multidrug-resistant P. aeruginosa from clinical isolates steadily increased in this secondary hospital in Korea in 2010. This study provides information about the antimicrobial-resistant patterns and genotype of multidrug-resistant P. aeruginosa isolated from clinical isolates in Korea, 2010.

          • SCIESCOPUSKCI등재

            Genomic Relationship of Salmonella enterica Serovar Typhimurium DT104 Isolates from Korea and the United States

            Kim,,Shukho,Chun,,Sung-Guen,Lim,,Ok-Young,Park,,Mi-Sun,Kang,,Yeon-Ho,Park,,Yong-Ho,Lee,,Bok-Kwon The Microbiological Society of Korea 2004 The journal of microbiology Vol.42 No.1

            Salmonella enterica serovar Typhimurium DT104 (Salmonella Typhimurium DT104 or DT104) has been emerging as a common pathogen for human in Korea since 1997. In order to compare the genomic relationship and to search for the dominant strains in Korea, we conducted pulsed-field gel electrophoresis (PFGE) and IS200 fingerprinting of 25 epidemiological unrelated isolates from human and animals from Korea and cattle from America. Two Salmonella Typhimurium DT104 isolates from human in Korea and all 8 isolates from American cattle had indistinguishable patterns from the PFGE and IS200 fingerprinting but multidrug-resistant Salmonella Typhimurium isolates, including DT104, from Korean animals had diverse genetic patterns. The data suggest that a dominant DT104 strain might have circulated between Korean and American cattle and that it had a high level of clonality.

          • SCIESCOPUSKCI등재

            Outbreaks of Imipenem-Resistant Acinetobacter baumannii Producing Carbapenemases in Korea

            Jeong,Seok-Hoon,Bae,Il-Kwon,Park,Kwang-Ok,An,Young-Jun,Sohn,Seung-Ghyu,Jang,Seon-Ju,Sung,Kwang-Hoon,Yang,Ki-Suk,Lee,Kyung-Won,Young,Dong-Eun,Lee,Sang-Hee The Microbiological Society of Korea 2006 The journal of microbiology Vol.44 No.4

            Among 53 Acinetobacter baumannii isolates collected in 2004, nine imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Korea. Nine carbapenemase-producing isolates were further investigated in order to determine the mechanisms underlying resistance. These isolates were then analyzed via antibiotic susceptibility testing, microbiological tests of carbapenemase activity, pI determination, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. One outbreak involved seven cases of infection by A. baumannii producing OXA-23 ${\beta}-lactamase$, and was found to have been caused by a single ERIC-PCR clone. During the study period, the other outbreak involved two cases of infection by A. baumannii producing IMP-1 ${\beta}-lactamase$. The two clones, one from each of the outbreaks, were characterized via a modified cloverleaf synergy test and an EDTA-disk synergy test. The isoelectric focusing of the crude bacterial extracts detected nitrocefin-positive bands with pI values of 6.65 (OXA-23) and 9.0 (IMP-1). The PCR amplification and characterization of the amplicons via direct sequencing showed that the clonal isolates harbored $bla_{IMP-1}$ or $bla_{oxA-23}$ determinants. The two clones were characterized by a multidrug resistance phenotype that remained unaltered throughout the outbreak. This resistance encompassed penicillins, extended-spectrum cephalosporins, carbapenems, monobactams, and aminoglycosides. These results appear to show that the imipenem resistance observed among nine Korean A. baumannii isolates could be attributed to the spread of an IMP-lor OXA-23-producing clone. Our microbiological test of carbapenemase activity is a simple method for the screening of clinical isolates producing class D carbapenemase and/or class B $metallo-{\beta}-lactamase$, in order both to determine their clinical impact and to prevent further spread.

          • SCOPUSKCI등재

            Diagnosis of Clostridium difficile infection in patients with hospital-acquired diarrhea

            Ibrahim,Afifi,,Salwa,Selim,Gomaa,,Fatma,Alzahraa,M.,Fathi,,Lamia,Fouad,Rasslan,,Fatma,Salah,Hamdy,,Ahmed,Mohamed The Microbiological Society of Korea 2018 미생물학회지 Vol.54 No.3

            Clostridium difficile infection (CDI) is a rapidly emerging infection that may have devastating consequences. Prompt and accurate diagnosis is crucial for management and control. The aim of this study was to determine the incidence of C. difficile associated diarrhea among hospitalized patients, and to compare different diagnostic laboratory methods for detection of toxin producing strains in clinical specimens. The study was conducted at a university hospital in Cairo during the period from May 2013 till June 2015. Subjects were under antibiotic therapy and presented with hospital-acquired diarrhea. Four hundred and sixty-five stool specimens were processed by different microbiological methods. C. difficile was recovered in culture in 51 of stool specimens. Of these, 86.3% to 98% were positive for toxin production by 2 different methods. This study showed that antibiotic intake is the major risk factor for development of hospital-acquired diarrhea. We evaluated different microbiological methods for diagnosis of C. difficile. We recommend the use of toxigenic culture as a gold standard for microbiological diagnosis of C. difficile.

          • KCI등재

            Microbiological, Physicochemical, and Antioxidant Properties of Plain Yogurt and Soy Yogurt

            임성미,Lim,,Sung-Mee The Microbiological Society of Korea 2013 미생물학회지 Vol.49 No.4

            양배추 피클로부터 분리된 유산균으로 발효하여 제조한 플레인 요구르트와 소이 요구르트를 저장하는 동안 미생물학적 및 물리화학적 변화와 항산화 특성을 조사하였다. 분리 균주는 당분해능과 유전자 염기서열 분석을 통해 L. casei PC05와 L. acidophilus PC16으로 동정되었다. Lactobacillus acidophilus PC16 균주로 발효시킨 요구르트의 적정산도, 점도 및 균수는 L. casei PC05 균주에 의해 제조된 요구르트에 비해 높게 나타났으며, 특히 플레인 요구르트보다 소이 요구르트에서 세균수와 물리화학적 요인의 측정값이 유의하게 높은 것으로 확인되었다. 한편, L. acidophilus PC16 균주로 발효시킨 요구르트는 DPPH 라디칼 소거능과 철이온 킬레이팅 활성이 높았으나, L. casei PC05 균주의 요구르트는 superoxide anion 제거능과 SOD 활성이 높은 것으로 나타났다. 대부분의 항산화능은 플레인 요구르트보다 소이 요구르트에서 더 높게 나타났으며, 요구르트의 지질과산화 억제능은 항산화 활성에 기인하는 것으로 추정된다. 또한 발효직후 요구르트의 미생물학적, 물리화학적 및 항산화 활성은 $4^{\circ}C$에서 8일간 저장하는 동안 거의 일정한 수준으로 유지되었다. This study evaluated the physicochemical and microbiological characteristics and antioxidant properties of yogurt samples fermented with lactic acid bacteria (LAB) obtained from pickled cabbage. API 50 CHL systems and 16S rRNA nucleotide sequence analyses revealed that the isolates were Lactobacillus casei PC05 and L. acidophilus PC16. Cell counts, titratable acidity, and viscosity of the yogurt samples fermented with L. acidophilus PC16 were significantly higher than those of the samples fermented with L. casei PC05 (P<0.05). The detected cell counts and physicochemical properties were significantly lower in plain yogurt than in soy yogurt (P<0.05). Yogurt samples fermented with L. acidophilus PC16 exhibited higher antioxidant activity, measured as ability to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and chelate ferrous ions, than those fermented with L. casei PC05. However, the ability to scavenge superoxide anions and superoxide dismutase (SOD) activity were significantly (P<0.05) higher in yogurt samples fermented with L. casei PC05 compared to those in samples fermented with L. acidophilus PC16. The antioxidant activity of soy yogurt was significantly (P<0.05) higher than that of plain yogurt. The antioxidant activity of the tested strains resulted in lipid peroxidation inhibition (in vitro), which may be related to the elimination of free radicals, chelating ability, and reducing power. There were no significant differences in the physicochemical properties and antioxidant activities of the yogurt samples during cold storage.

          • SCIESCOPUSKCI등재

            Purification and Characterization of Cold Active Lipase from Psychrotrophic Aeromonas sp. LPB 4

            Lee,,Han-Ki,Ahn,,Min-Jung,Kwak,,Sung-Ho,Song,,Won-Ho,Jeong,,Byeong-Chul The Microbiological Society of Korea 2003 The journal of microbiology Vol.41 No.1

            A lipase from Aeromonas sp. LPB 4, a psychrotophile isolated from a sea sediment was purified and characterized. The lipase was purified 53.5 fold to a homogeneous state by acetone precipitation and QAE sephadex column chromatography and its molecular weight was determined to be 50 kDa by SDS-PAGE. The enzyme exhibited maximum activity at 10$^{\circ}C$ and was stable at temperatures lower than 50$^{\circ}C$. This lipase favored substrates containing medium carbon chain of acyl group, while too low and high carbon chain decreased its activity. The lipolytic activity of purified lipase was slightly increased by the addition of 0.1% detergent, but decreased by 1% of detergent. Butanol severely decreased the lipase activity while methanol increased the activity about 15%.

          • SCOPUSKCI등재

            Cloning and Expression of the Extracellular $\beta$-lactamase gene from streptomyces sp. SMF13 in streptomyces lividans

            Rak,,Choi-Sang,Lee,,Kye-Joon The Microbiological Society of Korea 1992 미생물학회지 Vol.30 No.3

            Cloning of the gene encoding extracellular .betha.-lactamase from Streptomyces sp. SMF13 in a plasmid pIJ702 and expression of the gene in Streptomyces invidans were carried out. Optimal conditions for the formation of protoplasts of S.lividans and the regeneration of the protoplasts were evaluated. Streptomyces sp. SMF-13 was selected as a donor strain of .betha.-lactamase gene and totla DNA of the strain was partially digested with Sau3A I. DNA fragments ranged from 4kb to 10 kb were ligated to pIJ702 AT Bgl II site and then the ligated DNAs were transformed to the protoplasts of S, livivans. The transformation efficiency was $2 *10^{3}$ .$\mu$g DNA for the ligated DNA mixture. One colony among a thousand colonies regenerated showed extracellular .betha.-lactamase and the size of the inserted DNA fragment was estimated to be 3.94 kb. The .betha.-lactamase activity in the culture broth of the recombinant strain was maximum at 3 days culture to be 1.0 unit/ml.

          • SCOPUSKCI등재

            Transduction of the Wild-type polA Gene of Escherichia coli K-12 in a ColE1-Derived Mini-Mu Plasmid

            Parduez,,Nagy-Gyorgy,Choi,,Yong-Keel,Chung,,Young-Sup The Microbiological Society of Korea 1992 미생물학회지 Vol.30 No.2

            Teh $polA^{+}$ gene can be transducted in a multicopy mini-Mu plasmid, but not cloned because the product of this gene is lethal when overproduced. Although, we obtained one surviving cell, in which the ColEl-derived mini-Mu plasmid suffered a spontaneous deletion exactly at the region where the $polA^{+}$ gene was cloned. The $PolA^{+}$ unstream flanking sequence containing the promoter and pribnow-box was delected in vivo ; consequently this gene is not able to be expressed.

          • SCOPUSKCI등재

            Expression and Secretion of Heterologous Protein in Yeast

            Kim,,Moo-Kyum,Song,,Moo-Young,Yu,,Myeong-Hee,Yu,,Myeong-Hee,Park,,Hee-Moon,Kim,,Jinmi The Microbiological Society of Korea 1992 미생물학회지 Vol.30 No.2

            To investigate the expression and the secretion of heterologous proteins in yeast, we constructed an yeast secretion vector and produced a human secretory protein, .alpha.-1-antitrypsin (.alpha.-1-AT), from yeast cells. The secretion vector pGAT8 was constructed by inserting the signal sequence of yeast acid phosphatase gene (PH05) into the .alpha.1-AT expression vector pGAT6 which contained .alpha.-1-AT cDNA fused to GAL10-CYC1 promotor. The .alpha.-1-AT was produced efficiently in the yeast cells transformed with plasmid pGAT8, which was onfirmed both by the .alpha.-1-AT activity assay and by the immunoblot method using .alpha.-1-AT antibody. We also showed the secretion of .alpha.-1-AT into the culture media and into the periplasmic space by immunoblot.

          • SCOPUSKCI등재

            Cloning, Sequencing and Expression of an Extracellular Protease Gene from Serratia marcescens RH1 in Escherichia coli

            Lee,,Seung-Hwan,Kim,,Jeong-Min,Kwon,,Young-Tae,Kho,,Young-Hee,Rho,,Hyune-Mo The Microbiological Society of Korea 1992 미생물학회지 Vol.30 No.6

            Serratia marecescens RH1 isolated from soil samples produced large amount of extracellular proteases. One of the genes encoding an extracellular protease form S. marcescens RH1 was cloned in Escherichia coli by shot gun cloning method. The cloned protease, SSP, was stably expressed by its own promoter and excreted into the extracellular medium from E. coli host (ORF) of 3.135 nucleotides corresponding to 1.045 amino acids (112 kDa). The nucleotide and deduced amino acid sequence of SSP showed high overall homology (88%) to one of the S. marcescens protease (27), but low homology to other serine protease families. The optimal pH and temperature of the enzyme were pH 9.0 and 45.deg.C respectively. The activity of protease was inhibited by phenylmethylsulfonyl fluoride (PMSF), which suggests that the enzyme is a serine protease.

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