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T : 목차정보

Journal of bacteriology and virology : JBV 14건

  1. [국내논문]   Antisense Technology를 이용한 탄저균 치사 독소 발현 억제  

    박철민 (중앙대학교 의과대학 미생물학교실 ) , 김미정 (중앙대학교 의과대학 미생물학교실 ) , 안장훈 (중앙대학교 의과대학 미생물학교실 ) , 김기정 (중앙대학교 의과대학 미생물학교실 ) , 김원용 (중앙대학교 의과대학 미생물학교실 ) , 정상인 (중앙대학교 의과대학 미생물학교실)
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 247 - 259 , 2004 , 1598-2467 ,

    초록

    Bacillus anthracis is the causative agent of anthrax primarily in animals and rarely in humans. B. anthracis producing 'anthrax toxin', however, could be a major agent of biological warfare. Anthrax toxin is produced from the pXO1 plasmid encoding the lethal toxin (LeTx) consisted of the protective antigen (PA) and the lethal factor (LF). In this study, we tested whether specific antisense oligonucleotide could inhibit the gene expression in B. anthracis. The antisense oligonucleotide was forced into bacterial cells either by lipofection or heat shock method. The expression of LeTx in B. anthracis was analyzed by the Western blot analysis and the MTT assay using to Raw 264.7 cells. The LeTx protein was purified and used for the production of specific antibodies. The expression of LeTx could be confirmed only in B. anthracis strains haboring pXO1 plasmid. B. anthracis treated with the antisense oligonucleotide through heat shock method markedly inhibited the production of PA. In the Western blot analysis, the expression of PA was inhibited from $25\;{\mu}M$ and was completely inhibited at $50\;{\mu}M$ of the antisense oligonucleotide. In the MTT assay, the cytotoxicity was reduced to 20% at $20\;{\mu}M$ of the antisense oligonucleotide. Above results suggest that the antisense technology would be applied for the research on gene function in B. anthracis.

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  2. [국내논문]   Enrichment of Low Abundance Proteins of Helicobacter pylori Strain 26695 by the Heparin Chromatography  

    Lee, Woo-Kon ; Kim, Mi-Hye ; Song, Jae-Young ; Kim, Sam-Churl ; Park, Jeong-Uck ; Baik, Seung-Chul ; Kang, Hyung-Lyun ; Park, Seong-Gyu ; Hwang, Hyang-Ran ; Bae, Dong-Won ; Youn, Hee-Shang ; Ko, Gyung-Hyuck ; Cho, Myung-Je
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 261 - 272 , 2004 , 1598-2467 ,

    초록

    Low-abundance cellular proteins normally invisible on the standard two-dimensional SDS-polyacrylamide gel electrophoresis (2-DE SDS-PAGE) map must be enriched appropriately in order to be visualized and identified in cells or tissues. We applied proteins of H. pylori strain 26695 to a immobilized heparin-affinity resin, which has an affinity for nucleic acid-binding proteins, protein biosynthesis factors, and growth factors. The whole cell extract of H. pylori strain 26695 was fractionated by the heparin-agarose chromatography, and was analyzed by 2-DE. The 2-DE SDS-PAGE displayed spots after silver staining, which were identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). Among the ca. 150 spots that were processed, 79 proteins representing 57 genes were identified. Eleven proteins were determined to be nucleic acid-associated. Eighteen proteins were newly identified in this study, including DNA topoisomerase I. These results may provide guidance for enriching low abundance proteins of H. pylori and contribute to the construction of a master protein map of H. pylori.

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  3. [국내논문]   결핵균의 TSP 균체항원으로부터 특이항원의 분리 및 특성 분석   피인용횟수: 1

    백태현 (건양대학교 의과대학 미생물학교실 ) , 권혜숙 (건양대학교 의과대학 미생물학교실 ) , 이선 (건양대학교 의과대학 미생물학교실 ) , 이지숙 (건양대학교 의과대학 미생물학교실 ) , 조은경 (충남대학교 의과대학 미생물학교실 ) , 김화중 (충남대학교 의과대학 미생물학교실 ) , 이미리나 (건양대학교 의과대학 미생물학교실 ) , 유영춘 (건양대학교 의과대학 미생물학교실 ) , 박정규 (충남대학교 의과대학 미생물학교실)
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 273 - 282 , 2004 , 1598-2467 ,

    초록

    Tremendous efforts have been made to develop better vaccines and diagnostic markers for the effective control of tuberculosis. Recently, we reported that the Triton X-100 soluble protein (TSP) of Mycobacterium tuberculosis induced strong T-cell proliferation and IFN- $\gamma$ production in humans, and also conferred a significant level of protection against tuberculosis in a mouse model. In this study, the TSP was prepared by Triton X-100 extraction of Mycobacterium tuberculosis bacilli, which was followed by Triton X-114 phase partitioning. Western blot analysis using sera of 177 active pulmonary tuberculosis patients and 323 healthy individuals revealed that the TSP contained a immunodominant 40-kDa antigen specifically reacting with some sera from pulmonary tuberculosis patients. The 40-kDa antigen was purified by ion-exchange chromatography, and partially characterized by two-dimensional gel electrophoresis and N-terminal sequencing. Results of this study suggest that 40-kDa molecule of the TSP antigen from the cell surface of Mycobacterium tuberculosis can be used as a serodiagnostic marker as well as a potential vaccine candidate against tuberculosis.

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  4. [국내논문]   Detergent로 추출한 Orientia tsutsugamushi 세포외막 단백질의 세포 부착  

    김미경 (인하대학교 의과대학 미생물학교실 ) , 김미정 (인하대학교 의과대학 미생물학교실 ) , 임병욱 (인하대학교 의과대학 미생물학교실 ) , 강재승 (인하대학교 의과대학 미생물학교실)
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 283 - 289 , 2004 , 1598-2467 ,

    초록

    Orientia tsutsugamushi, a causative agent of scrub typhus, is an obligate intracellular parasite. The mechanisms by which O. tsutsugamushi invade host cells are unknown. Given the importance of surface-exposed proteins in the pathogenesis of microbial pathogens, outer membrane proteins (OMP) of O. tsutsugamushi were extracted with detergents and their cellular binding was studied. Outer membrane fraction of O. tsutsugamushi was enriched by a sodium-lauryl sarcosinate (Sarkosyl) treatment of total membranes. Outer membrane proteins were extracted by the treatment with sodium dodecyl sulfate (SDS) and Sarkosyl. The resulting soluble fractions were examined for their cellular binding by the immunofluorescence microscopy. A fifty six kilodalton protein was found to bind to fixed ECV304 cells only when the outer membrane preparation was not treated by DTT or heat. These results suggest that the conformation the 56-kDa OMP is important for the attachment to the host cell surfaces.

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  5. [국내논문]   A Two Component Signal Transduction System Required for the Virulence of Vibrio vulnificus  

    Jin, Yeon-Soo ; Kim, Young-Ran ; Rhee, Joon-Haeng
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 291 - 301 , 2004 , 1598-2467 ,

    초록

    Vibrio vulnificus is an estuarine bacterium that opportunistically infects humans with underlying hepatic diseases, heavy alcohol drinking habits, and other immunocompromised conditions. A locus in the V. vulnificus genome was cloned and sequenced, which showed similarities to the bacterial two-component signal transduction system. The locus encoded a putative sensor kinase, designated vvgS, and a divergently transcribed putative response regulator, designated vvgR. VvgS was predicted to be a protein belonging to the tripartite hybrid sensor kinase subfamily such as ArcB and BvgS. VvgR showed similarities to well known response regulators. An insertional mutation of vvgS in a V. vulnificus strain led to a remarkable decrease of cytotoxicity to HeLa cells, while the production of exotoxins, the hemolysin and the protease, slightly increased by the vvgS mutation. Adhesion to HeLa cells only slightly decreased. However, the vvgS mutation had no effect on the motility of V. vulnificus. The vvgS mutation resulted in a significant decrease of lethality to mice. These results indicate that vvgRS two-component system plays a very important role in regulating novel virulence factor(s) of V. vulnificus associated with cytotoxicity other than hemolysin and protease during the infection process.

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  6. [국내논문]   Prevalence of CTX-M Extended-spectrum Beta-lactamases in Clinical Isolates of Enterobacteriaceae in Korea   피인용횟수: 1

    Kim, Jung-Min ; Lim, Yu-Mi
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 303 - 310 , 2004 , 1598-2467 ,

    초록

    The evolution and dissemination of extended-spectrum $\beta$ -lactamases (ESBL) have compromised the clinical use of third-generation cephalosporins worldwide. Although most ESBLs belong to the TEM and SHV $\beta$ -lactamase families, the members of CTX-M, a novel ESBL family, are increasing worldwide in Gram-negative bacteria. We examined the prevalence of CTX-M ESBL in clinical isolates of the family Enterobacteriaceae collected from three university hospitals located in three different cities in Korea. Among a total of 603 isolates collected, 163 isolates (27.0%) revealed ${\geq}2\;{\mu}g/ml$ of MIC against cefotaxime, and 93 isolates (15.4%) produced ESBL confirmed by the double disk synergy test. Among 93 ESBL-producing isolates, $bla_{CTX-M}$ genes were detected in 41 isolates by PCR method and they included 1 isolate of C. freundii, 3 of E. aerogenes, 2 of E. cloacae, 17 of E. coli, 9 of K. pneumoniae, and 9 of S. marcescens. Thus, the overall prevalence of CTX-M ESBL-producing isolates among the family Enterobacteriaceae was 6.8% (41 of 603 isolates) and the proportion of CTX-M-producers among the ESBL-producing isolates was 44.1 % (41 of 93 isolates). Further determination of the $bla_{CTX-M}$ subtype by nucleotide sequencing revealed $bla_{CTX-M-3}$ in 17, $bla_{CTX-M-15}$ in 11, $bla_{CTX-M-14}$ in 9, and $bla_{CTX-M-9}$ in 4 isolates. To our knowledge, this is the first report on the dissemination of CTX-M ESBL among clinical isolates of the family Enterobacteriaceae in Korea.

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  7. [국내논문]   Differential Mast Cell Response to Different Bacteria  

    Kim, Jug-Min ; Abraham, Soman N.
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 311 - 319 , 2004 , 1598-2467 ,

    초록

    Mast cells (MCs) are highly specialized for the synthesis and secretion of pharmacologically active products. Although implicated in various inflammatory diseases such as asthma, allergy, multiple sclerosis, and Crohn's disease, MCs have also an important physiologic role in immunosurveillance and modulation of host's innate immune responses following bacterial infection. Here, we present that MCs show varying capability of recognizing and responding to different bacteria. Whereas MCs were readily degranulated and released neutrophil chemoattractants in response to E. coli or S. aureus infections, they failed to degranulate during S. typhimurium infections. Consequently, E. coli infections were associated with a vigorous neutrophil response and early bacterial clearance whereas S. typhimurium infections were associated limited neutrophil influx and bacterial multiplication at sites of infection. Interestingly, injection of compound 48/80, a MC specific activator, at sites of S. typhimurium infection triggered MC mediated neutrophil influx and bacterial clearance.

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  8. [국내논문]   The Synergistic Effect of LMP1 and IRF7 on the Expression of TAP1 in DG 75 Cell Line  

    Park, Ho-Sun
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 321 - 329 , 2004 , 1598-2467 ,

    초록

    In this study, the author explored the role of interferon regulatory factor 7 (IRF7) and latent membrane protein 1 (LMP1) on the regulation of antigen presenting molecules in B-lymphoblastoid cell lines. First, the author observed the endogenous expression of IRF7 and LMP1 in paired EBV-positive B-lymphoblastoid cell lines, Sav I and Sav III, which represent EBV type I and type III latency, respectively. The Sav I cell, which does not express LMP1, showed very low levels of endogenous IRF7 in the cytoplasm. However, Sav III cells, which express large amounts of LMP1, contained high levels of endogenous IRF7 in both the cytoplasm and the nucleus. The expression of surface MHC class I antigen was 7.8-fold higher in Sav III compared with Sav I cells when measured by fluorescence activated cell sorter (FACS) analysis. To understand whether IRF7 is involved in regulation of MHC I and TAP1, LMP1 or IRF7 were expressed by cotransfection in DG75 cells. Levels of TAP1 protein were up-regulated by LMP1 and IRF7 alone, and by LMP1 co expression with IRF7, the expression level was highest after co-transfection of LMP1 with IRF7. TAP1 promoter activity was also up-regulated to 2.4, 2.0, 3.2-fold by LMP1, by IRF7, and by LMP1 plus IRF7, respectively. Surface expression of MHC class I antigen was up-regulated by LMP1 alone and LMP1 plus IRF7, but not by IRF7 alone. These results suggest that IRF7 induces the expression of TAP1, but not MHC class I antigen and that LMP1 and IRF7 have additive effects on the expression of TAP1 protein.

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  9. [국내논문]   Detection of Enteroviruses and Mammalian Reoviruses by RT-PCR and Integrated Cell Culture-PCR in CPE-positive Surface Water Samples  

    Kim, Hee-Jung ; Ko, Hyun-Ae ; Kim, Sang-Hyun
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 331 - 338 , 2004 , 1598-2467 ,

    초록

    The environmental water samples assayed by total culturable virus assay (TCVA) were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and integrated cell culture-PCR (ICC-PCR) method for enteroviruses and reovirus since they are the usually detected virus groups by culture assays. The detection sensitivities of the TCVA, RT-PCR, and ICC-PCR were compared and the overall reliability of the detection was analyzed to confirm environmental samples for enteric viruses. A total of eight samples from different areas was analyzed by performing TCVA, RT-PCR, and ICC-PCR. Virus concentrations in surface water samples ranged from 1.03 to 47.3 most probable numbers of infectious units (MPN) per 100 liters. When primers specific for both enteroviruses and reoviruses were used in both RT-PCR and ICC-PCR, all the samples (100%) were positive for the viruses. Reoviruses were the most frequently detected ones among the samples. According to the sequence results of enteroviruses, five of the samples were contaminated by coxsackievirus type B3, and the rest by coxsackievirus type B6, echovirus type 30, or vaccine strain poliovirus type 3. It was observed that both enteroviruses and reoviruses were detected concurrently in all CPE-positive environmental water samples.

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  10. [국내논문]   낮은 복제수 플라스미드를 활용한 일본뇌염바이러스 SA14-14-2주의 Full-Length cDNA 클론의 제작 및 안정적 유지  

    민경일 (식품의약품안전청 생물의약품평가부 백신과 ) , 김영민 (식품의약품안전청 생물의약품평가부 백신과 ) , 추미성 (식품의약품안전청 생물의약품평가부 백신과 ) , 백선영 (식품의약품안전청 생물의약품평가부 백신과 ) , 김재옥 (식품의약품안전청 생물의약품평가부 백신과 ) , 류승렬 (식품의약품안전청 생물의약품평가부 백신과 ) , 민복순 (식품의약품안전청 생물의약품평가부 백신과 ) , 김연희 (식품의약품안전청 생물의약품평가부 백신과 ) , 박미경 (식품의약품안전청 생물의약품평가부 백신과 ) , 변우현 (강원대학교 자연과학대학 미생물학과 ) , 허숙진 (식품의약품안전청 생물의약품평가부 백신과)
    Journal of bacteriology and virology : JBV v.34 no.4 ,pp. 339 - 353 , 2004 , 1598-2467 ,

    초록

    Recently the reverse genetics system contributed to the progresses in the investigation of positive-stranded RNA viruses. Here, we report the successful construction of a stable full-length infectious cDNA clone of the live attenuated JEV vaccine strain SA14-14-2. The eleven kilobase viral RNA genome was reverse transcribed, amplified as four overlapping DNA fragments and successively ligated into the low copy number plasmid pACYC184, which contains the pl5A origin of replication. In vitro-transcribed RNAs had a specific infectivity of approximately $10^4\;PFU/{\mu}g$ RNA, and the resulting virus exhibited growth kinetics and plaque morphology similar to the parental virus in cell culture. The structural and functional integrity of the cDNA clone was stably maintained even after at least 150 generations in Escherichia coli strain TOP10. The cDNA clone was engineered to contain single nucleotide change to create a XhoI site and knock out a XbaI site (A to C at nt 9134) acting as a genetic marker. This genetic marker was retained in the recovered progeny virus. Our results suggest that the instability of the full-length infectious JEV cDNA clone can be overcome by employing low copy number plasmid pACYC184. This infectious JEV cDNA clone will aid future studies of pathogenesis, virulence, and replication. Furthermore, it will facilitate the development of SA14-14-2 based recombinant vaccines.

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