• Title/Summary/Keyword: typhimurium

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세포내 기생세균의 병원성 관련 유전자의 분석에 관하여 (Analysis of Genes Involved in the Pathogenesis of Intracellularly Survival Bacteria)

  • 전태일;이태윤;김성광
    • Journal of Yeungnam Medical Science
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    • 제9권2호
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    • pp.248-255
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    • 1992
  • S. typhimurium의 세포내생존을 가능케하는 유전자가 다른 세균에서도 존재하는지를 조사하기 위해 8주의 세균주를 사용하였다. phoP-PhoQ operon은 세포내 환경의 자극을 인지하고 그 환경의 적응에 관여하는 유전자의 유전자표현을 조절한다고 알려져 있다. S. typhimurium의 phoP region의 514 basepairs EcoRV DNA fragment를 probe로 dot blot hybridization을 실시하였다. K. pneumoniae, P. aeruginosa, S. marscescens, E. cloacae, S. typhimurium의 phoP/phoQ gene과 유사한 DNA sequence를 가지지 않았으며 E. coli, S. dysenteriae, E. cloacae에서는 세포내 생존가능세균이 아님에도 불구하고 positive signal을 나타내었다. 이상의 결과에서 S. typhimurium외의 세포내 생존가능세균에는 phoP/PhoQ operon이 없다는 것을 알게 되었고 세포내 생존이 가능하지는 않지만 S. typhimurium과 계통발생학적으로 가까운 세균주에서 phoP/phoQ operon이 발견되었다. L. monocytogenes의 세포내 생존에는 phoP/phoQ에 의존하지 않는 어떤 다론 기전이 존재할 것으로 사료된다.

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송화 메탄올 추출물의 항산화적 항돌연변이 효과 (Antimutagenic and Antioxidative Effects of Methanol Extract of Pine Pollen)

  • 박정섭;안병용;최동성
    • 한국식품영양학회지
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    • 제16권4호
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    • pp.303-309
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    • 2003
  • 직ㆍ간법 변이원과 산화적 돌연변이원에 대한 송화분 추출물의 항돌연변이원성에 대하여 Ames test로 조사하였다. 메탄을 추출물을 이용하여, 10종의 돌연변이에 대한 돌연변이 억제효과를 검색한 결과 S. typhimurium TA98에서는 daunomycin, AFB$_1$과 Trp-P-1의 변이원성에 대해서만 각각 17.8, 82.2, 80.9%의 돌연변이 억제효과를, S. typhimurium TA100에서는 AFB$_1$의 변이원성에 대해서만 72.3%의 돌연변이 억제효과를 나타내었으며, S. typhimurium TA102, 104에서 t-과산화부틸과 $H_2O$$_2$의 돌연변이원성에 대한 메탄을 추출물의 돌연변이 억제 효과는 t-과산화부틸을 돌연변이원으로 한 S. typhimurium TA102에서 16.3%의 억제효과를 선택적으로 나타내었다. 따라서 메탄을 추출물을 클로르포름, 에틸아세테이트, 부탄올, 물 순으로 순차용매 분리한 다음, 이 분획물에 대한 항돌연변이원성을 검색하였다. 그 결과 S. typhimurium TA98에서 daunomycin, AFB$_1$과 Trp-P-1에 대해서 플로로포름 분획물은 각각 55.6, 93.7, 93.5%, 에틸아세테이트 분획물은 각각 11.4, 74.3, 85.2%, 부탄을 분획물에서는 10.9, -5.6, 6.8%, 물 분획물은 각각 -3.4, -3.8, 4.6%의 돌연변이 억제효과를 나타내었다. S. typhimurium TA100에서는 AFB$_1$에 대해 클로로포름 분획물과 에틸아세테이트 분획물이 각각 95.1, 62.5%의 억제효과를 나타내었으며, S. typhimurium TA102에서는 t-과산화부틸에 의해 유도된 산화적 돌연변이에 대해 클로로포름 분획물이 93.6%로 매우 강한 억제효과를 나타내었으며, 기타 분획물에서는 매우 낮은 억제효과를 나타내었다.

육회와 육사시미에 접종된 Salmonella Typhimurium와 Listeria monocytogenes 검출을 위한 Loop-mediated isothermal amplification와 식품공전의 배지 시험법, real-time PCR의 검출 성능 비교 (Comparison of Loop-mediated Isothermal Amplification and Korea Standard Food Codex (KFSC) Method for Detection of Salmonella Typhimurium, Listeria monocytogenes Artificially Inoculated in Yuk-hwe and Yuk-sashimi)

  • 곽승해;이소영;김진희;오세욱
    • 한국식품위생안전성학회지
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    • 제34권3호
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    • pp.277-282
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    • 2019
  • 본 연구에서는 한국 전통 식품에서 Salmonella Typhimurium와 Listeria monocytogenes의 검출에 대하여 LAMP에 기반한 3M Molecular Detection Assay 2 (3M MDA 2)와 식품공전에 등재된 분리배지, real-time PCR의 검출 성능을 비교하고자 하였다. 육회와 육사시미에 $10^0-10^4CFU/25g$의 수준으로 S. Typhimurium와 L. monocytogenes을 각각 접종하였다. Citrobacter freundii와 Listeria innocua는 S. Typhimurium와 L. monocytogene의 검출에 영향을 주는 균으로 사용하였다. S. Typhimurium와 L. monocytogenes만 $10^0-10^4CFU/25g$ 수준으로 접종한 모든 시료에서는 분리배지, real-time PCR과 LAMP에서 양성으로 검출되었다. C. freundii와 L. innocua를 같이 접종한 경우에서 부분적으로 양성이 나타났다. 육회와 육사시미에 대하여 real-time PCR 보다 3M MDA 2가 더 검출효율이 높음을 알 수 있었다. 분리배지가 가장 검출효율이 높았지만 3M MDA 2와 큰 차이가 없었다. 배지를 사용하는 방법은 최소 일주일의 시간이 소요되고 PCR의 경우 inhibitor의 영향을 많이 받아 정확한 검출이 어려운 점이 있다. 그러나 LAMP에 기반한 3M MDA 2는 enrichment 후 다음 날 간단한 protocol을 통해 25분 이내로 샘플의 양성 반응을 확인할 수 있어 식중독균에 대해 신속하고 정확한 검출이 가능한 것으로 판단되었다.

Genetic Diversity of Multi-resistant Salmonella enterica Serotype Typhimurium Isolates from Animals and Humans

  • Woo Yong-Ku;Lee Su-Hwa
    • Journal of Microbiology
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    • 제44권1호
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    • pp.106-112
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    • 2006
  • In this study, the genetic diversities of multi-resistant Salmonella typhimurium (ST) isolates were analyzed via the application of both pulsed field gel electrophoresis (PFGE) and Polymerase chain reaction (PCR) analysis methods, using 6 kinds of primers (REP, ERIC, SERE, BOX, P-1254 and OPB-17). And their discriminative abilities (DA) were also compared in order to determine the most effective and reliable analysis method. 118 S. typhimurium isolates, cultured from diverse animals and human patients in Korea beginning in 1993, were analyzed and subjected to a comparison of Simpson's index of diversity (SID), using both PFGE and PCR methods. PFGE by XbaI enzyme digestion allowed for discrimination into 9 pulsotypes, with high SID values (0.991) on the genomic DNA level. This shows that PFGE is a very discriminative genotypic tool, and also that multiple clones of S. typhimurium isolates had existed in domestic animals and humans in Korea since 1993. However, we could ultimately not to trace the definitive sources or animal reservoirs of specific S. typhimurium isolates examined in this study. Depending on the SID values, the combined method (7 kinds of method) was found to be the most discriminative method, followed by (in order) SERE-PCR, REP-PCR, ERIC-PCR, PFGE & OPB-17 (RAPD), P-1254 (RAPD), and BOX-PCR at the $80\%$ clone cut-off value. This finding suggests that the REP-PCR method (which utilizes 4 primer types) may be an alternative tool to PFGE for the genotyping of S. typhimurium isolates, with comparable cost, time, and labor requirement. The establishment of a highly reliable and discriminatory method for epidemiologic analysis is considered necessary in order for researchers to trace the sources of specific pathogens and, consequently, to control and prevent the spread of epidemic S. typhimurium isolates to humans.

Genomic Approaches for Understanding the Characteristics of Salmonella enterica subsp. enterica Serovar Typhimurium ST1120, Isolated from Swine Feces in Korea

  • Kim, Seongok;Kim, Eunsuk;Park, Soyeon;Hahn, Tae-Wook;Yoon, Hyunjin
    • Journal of Microbiology and Biotechnology
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    • 제27권11호
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    • pp.1983-1993
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    • 2017
  • Salmonella enterica subsp. enterica serovar Typhimurium, one of the most common foodborne pathogens, is transmitted mainly through contaminated food derived from infected animals. In this study, S. Typhimurium ST1120, an isolate from pig feces in Korea, was subjected to whole-genome analysis to understand its genomic features associated with virulence. The genome of ST1120 was found to have a circular chromosome of 4,855,001 bp (GC content 52.2%) and a plasmid of 6,863 bp (GC content 46.0%). This chromosome was predicted to have 4,558 open reading frames (ORFs), 17 pseudogenes, 22 rRNA genes, and 86 tRNA genes. Its plasmid was predicted to have three ORFs. Comparative genome analysis revealed that ST1120 was phylogenetically close to S. Typhimurium U288, a critical isolate in piggery farms and food chains in Europe. In silico functional analysis predicted that the ST1120 genome harbored multiple genes associated with virulence and stress resistance, including Salmonella pathogenicity islands (SPIs containing SPI-1 to SPI-5, SPI-13, and SPI-14), C63PI locus, ST104 prophage locus, and various antibiotic resistance genes. In accordance with these analysis results, ST1120 showed competence in invasion and survival abilities when it was added to host cells. It also exhibited robust resistance against antibiotics in comparison with other S. Typhimurium strains. This is the first report of the complete genome sequence of S. Typhimurium isolated from swine in Korea. Comparative genome analysis between ST1120 and other Salmonella strains would provide fruitful information toward understanding Salmonella host specificity and developing control measures against S. Typhimurium infection.

Multiplex PCR 기법을 이용한 Salmonella Enteritidis와 S. Typhimurium의 특이적 검출에 관한 연구 (Identification of Salmonella Enteritidis and S. Typhimurium by multiplex polymerase chain reaction)

  • 이우원;이승미;이강록;이동수;박호국
    • 한국동물위생학회지
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    • 제32권2호
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    • pp.147-153
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    • 2009
  • Salmonella species are the most important etiologic agents of food-borne acute gastroenteritis. The most common serotypes isolated from humans are Salmonella enterica serotype Typhimurium (S. Typhimurium) and S. Enteritidis. Traditional detection methods for Salmonella are based on cultures using selective media and characterization of suspicious colonies by biochemical and serological tests. These methods are generally time-consuming and not so highly sensitive. Recently, the polymerase chain reaction (PCR) has been used as a highly sensitive, specific, and rapid test for the presence of pathogenic bacteria. In this study, a multiplex PCR (m-PCR) was used to detect S. Typhimurium and S. Enteritidis. We selected m-PCR target genes, which were the spv (virulence plasmid specific for S. Enteritidis) and sefA (S. Enteritidis fimbrial antigen) genes, fliC (H1-i antigen specific for S. Typhimurium) and a randomly cloned sequence specific for the genus Salmonella. With m-PCR, random sequence was detected from all strains of Salmonella spp, spv and sefA were detected from all strains of S. Enteritidis (100%), and fliC was detected from all strains of S. Typhimurium (100%). This assay indicate that the specificity of the m-PCR make them potentially valuable tools for detection of S. Typhimurium and S. Enteritidis.

난백 유래 라이소자임의 마우스 살모넬라증에 대한 항균 및 치료 효과 (Evaluation of antibacterial and therapeutic effects of egg-white lysozyme against Salmonella Typhimurium in ICR mice infected with Salmonella Typhimurium)

  • 김희규;황재승;제우영;손송이;이후장
    • 대한수의학회지
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    • 제56권2호
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    • pp.103-108
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    • 2016
  • Salmonellosis is a major bacterial zoonosis that causes self-limited enteritis in animals and foodborne disease and typhoid fever in humans. Recently, multi-drug-resistant strains of Salmonella spp. have increased and caused more serious problems in public health. The present study investigated the antibacterial effects of egg-white lysozyme (EWL) against Salmonella (S.) Typhimurium and the therapeutic effects of EWL for murine salmonellosis. Evaluation of the antibacterial effects of EWL against S. Typhimurium revealed a minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of EWL of 6.25 and $300{\mu}g/mL$, respectively. In the bacterial growth inhibition test, EWL at 300 (p < 0.05) and $600{\mu}g/mL$ (p < 0.01) significantly inhibited the growth of S. Typhimurium at 4 h post-incubation. EWL administration at MIC (LYS-1), MBC (LYS-2) and $2{\times}MBC$ (LYS-3) for 14 days resulted in mortality of mice infected with S. Typhimurium of 70, 40 and 10%, respectively, while that of control mice (CON) was 90%. Counts of S. Typhimurium in murine spleens were significantly lower in LYS-2 and LYS-3 than CON (p < 0.05). The results of this study indicate that EWL has the potential for treatment of ICR mice infected with S. Typhimurium.

생육조건에 따른 Salmonella typhimurium의 숙주세포 부착성 (Effect of Growth Conditions in the Attachment of Salmonella typhimurium to the Host Cells)

  • 김영희;김상웅;강호영
    • 생명과학회지
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    • 제15권4호
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    • pp.641-646
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    • 2005
  • Salmonella typhimurium TML이 Int-407 숙주세포에 부착하는 정도는 S. typhimurium이 낮은 농도의 철이 함유된 배지에서 배양되었을 때보다 LB 액체배지 또는 높은 농도의 철이 함유된 배지에서 생육한 것에서 약 10배 정도의 높은 수준으로 관찰되었다. 고농도의 철이 포함된 배양조건이 살모넬라가 숙주세포에 부착시키는 정도를 향상시키는데 반해, 칼슘, 코발트, 구리, 인산, 마그네슘 그리고 망간과 같은 다른 양이온은 그렇지 않다. 이것은 아마도 철이 Salmonella가 부착에 필요한 요소들의 발현을 활성화하는 역할을 하는 것으로 추정된다. 철 농도에 따른 부착정도의 차이들은 type 1 fimbriae, mannose resistant hemagglutinin과 flagellum등을 생성하지 않는 다양한 S. typhimurium 돌연변이주들에서도 관찰되는 것으로 미루어보아 이들 구조체들과 상관이 없는 밝혀지지 않은 어떤 인자가 부착성 증가에 관여하는 것으로 사료된다. fur유전자가 불활성화된 S. typhimurium 돌연변이주의 부착성이 야생형 Salmonella와 유사한 방식으로 철에 의해 조절되었는데, 이는 철 농도에 따른 부착성의 변화에 관여하는 잠재적인자의 발현이 Fur 단백질과 독립적으로 이루어진다는 것을 나타낸다.

Targeting Orthotopic Glioma in Mice with Genetically Engineered Salmonella typhimurium

  • Wen, Min;Jung, Shin;Moon, Kyung-Sub;Jiang, Shen Nan;Li, Song-Yuan;Min, Jung-Joon
    • Journal of Korean Neurosurgical Society
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    • 제55권3호
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    • pp.131-135
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    • 2014
  • Objective : With the growing interests of bacteria as a targeting vector for cancer treatment, diverse genetically engineered Salmonella has been reported to be capable of targeting primary or metastatic tumor regions after intravenous injection into mouse tumor models. The purpose of this study was to investigate the capability of the genetically engineered Salmonella typhimurium (S. typhimurium) to access the glioma xenograft, which was monitored in mouse brain tumor models using optical bioluminescence imaging technique. Methods : U87 malignant glioma cells (U87-MG) stably transfected with firefly luciferase (Fluc) were implanted into BALB/cAnN nude mice by stereotactic injection into the striatum. After tumor formation, attenuated S. typhimurium expressing bacterial luciferase (Lux) was injected into the tail vein. Bioluminescence signals from transfected cells or bacteria were monitored using a cooled charge-coupled device camera to identify the tumor location or to trace the bacterial migration. Immunofluorescence staining was also performed in frozen sections of mouse glioma xenograft. Results : The injected S. typhimurium exclusively localized in the glioma xenograft region of U87-MG-bearing mouse. Immunofluorescence staining also demonstrated the accumulation of S. typhimurium in the brain tumors. Conclusion : The present study demonstrated that S. typhimurium can target glioma xenograft, and may provide a potentially therapeutic probe for glioma.

Identification of the Invasion Determinants of Salmonella typhimurium for Cultured HEp-2 and HeLa Cells

  • Park, Jeong-Uck;Joo, Woo-Hong
    • Journal of Life Science
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    • 제10권1호
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    • pp.6-9
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    • 2000
  • Salmonella typhimurium is a causative agent of the common worldwide disease, salmonellosis. To identify putative invasion genes involved in Samonella infections, a S. typhimurium cosmid library was constructed in noninvasive E. coli DJl. The invasion efficiencies of the cosmid library for cultured HEp-2 and HeLa cells were estimated by tissue-culture invasion assay. 2 out of 1,000 transductants, DHl(pSI623) and DHl(pSI511) were able to invade the cells. Compared to E. coli by DHl(pSI511) increased 25- and 33 fold, respectively. The invasion efficiencies of HeLa cells by DHl(pSI623) increased 31- and 35 fold, respectively. This illustrates that the cosmid clones, DHl(pSI623) and DHl(pSI511) could harbor the invasion determinants derived from genomic DNA of S. typhimurium 82/6915, conferring the invasive characters for the cells.