• 대한의생명과학회지
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• 제21권4호
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• pp.233-240
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• 2015

Some virulence proteins of Helicobacter pylori, such as vacuolating cytotoxic protein A (VacA) and cytotoxin-associated gene protein A (CagA) have been reported to be causative agents of various gastric diseases including chronic gastritis, gastric ulcer or gastric adenocarcinoma. The expression level of these virulence proteins can be regulated when H. pylori is exposed to the antibacterial agent, cyanidin 3-O-glucoside (C3G) as previously reported. In this study, we analyzed the quantitative change of various virulence proteins including CagA and VacA by C3G treatment. We used 2-dimensional electrophoresis (2-DE) to analyze the quantitative change of representative ten proteome components of H. pylori 60190 ($VacA^+/CagA^+$; standard strain of Eastern type). After 2-DE analysis, spot intensities were analyzed using ImageMaster$^{TM}$ 2-DE Platinum software then each spot was identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) or peptide sequencing using Finnigan LCQ ion trap mass spectrometer (LC-MS/MS). Next, we selected major virulence proteins of H. pylori among quantitatively meaningful ten spots and confirmed the 2-DE results by Western blot analysis. These results suggest that cyanidin 3-O-glucoside can modulate a variety of H. pylori pathogenic determinants.

• 대한의생명과학회지
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• 제27권4호
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• pp.195-207
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• 2021

Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterial pathogen capable of causing human diseases, such as soft tissue infection, bacteremia, endocarditis, toxic shock syndrome, pneumonia, and sepsis. Although the incidence rate of diseases caused by MRSA has declined in recent years, these diseases still pose a clinical threat due to their consistently high morbidity and mortality rates. However, the role of virulence factors in staphylococcal infections remains incompletely understood. Methicillin resistance, which confers resistance to all β-lactam antibiotics in cellular islets, is mediated by the mecA gene in the staphylococcal cassette chromosome mec (SCCmec). Differences in SCCmec types and differences in their sizes and structures serve epidemiological purposes and are used to differentiate between hospital-associated (HA)-MRSA and community-associated (CA)-MRSA. Some virulence factors of S. aureus are also providing a distinction between HA-MRSA and CA-MRSA. These factors vary depending on the presence of toxins, adhesion, immune evasion, and other virulence determinants. In this review, we summarized an overview of MRSA such as resistance mechanisms, SCCmec types, HA- and CA-MRSA, and virulence factors that enhance pathogenicity or MRSA epidemiology, transmission, and genetic diversity.

• 한국동물위생학회지
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• 제32권1호
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• pp.61-76
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• 2009

At the present study, it was aimed to explore the molecular genetic characterization of multiple antimicrobial resistant Salmonella spp. isolates from pigs and cattle. A total of 138 Salmonella Typhimurium (S. Typhimurium) isolates were typed with phage, among them, 83.3% of S. Typhimurium tested could divide into a 10 phage types. Definitive type 193 (DT193) (25.4%) and DT195 (24.6%) were exhibited as the dominant types. DT104 and U302 were found from pigs and cattle. On the other hand, S. Enteritidis had 6 phage types, of them, phage type 21 (PT21) and PT11b were the popular types. In the plasmid profiles, 135 of S. Typhimurium isolates were exhibited 1 to 6 plasmid bands which molecular weight ranged from 90 to 2kb. 35 isolates (25.4%) harbored a 90kb plasmid which is thought to be the serotype specific virulence plasmid. Two of twenty five S. Enteritidis had common plasmids at 2 and 1.5kb. With multiplex polymerase chain reaction, virulence genes (invA and spvC) were detected from all Salmonella spp. from 167 of S. Typhimurium, S. Enteritidis and chloramphenicol resistant S. Schwarzengrund, but some drug resistant genes, such as PSE-1, cml/tetR and flo were not determined but other drug resistant genes, for example TEM and int were found. The detection rates of spvC, TEM and int gene was 35.3%, 29.3% and 72.5%, respectively. The TEM gene was highly popular in S. Typhimurium, which was detected from ampicillin and amoxicillin resistant strains as 95.9%. int gene was able to detect from all the isolates identified as multidrug resistsnt (MDR), particularly DT193 was thought as the most prevalent virulence and multidrug resistance isolate. The major plasmid profile and drug resistance pattern of DT193 were 90, 40, 10.5, 6.3, 3.0kb and ACCbDNaPSSuT, respectively. MDR was commonly found in other phage types, particularly DT104, U302 and DT203.

• 한국식품위생안전성학회지
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• 제32권2호
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• pp.135-140
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• 2017

본 연구는 서울시내에서 시판중인 식육에서 E. faecalis를 분리하고 이 균들의 항생제 내성 패턴, 항생제 유출 펌프 유전자 및 병독성 유전자의 분포를 분석하였다. 총 277개의 식육시료에서 93균주의 E. faecalis 를 분리하였다. 이 균주들의 항생제 내성비율은 ampicillin에는 35.5%, chloramphenicol에 6.4%, ciprofloxacin에 4.3%, eryhtromycin에 18.3%, quinupristin-dalfopristin에 76.3%, tetracycline에 45.2%의 내성이었으며 levofloxacin, teiconplanin 및 vancomycin에는 모든 균이 감수성이었다. 약물 유출펌프인 MFS 타입의 eme(A)와 ABC 타입의 efr(A)유전자는 모든 균주(100%)에서 확인되었으며 efr(B)는 98.9%, lsa는 91.4%의 균주에서 확인되었다. 병독성 인자인 gel(E)는 68.8%, ace는 90.3%, asa1는 47.3%, efaA는 91.4%, esp는 12.9%의 균주에서 확인되었다. 본 연구는 시판 식육에서 분리한 지표 미생물의 하나인 E. faecalis의 항생제 내성, 약물유출 펌프 및 병독서 유전자의 분포를 분석한 연구로 지속적인 모니터링을 하여야 할 것으로 사료된다.

• 대한미생물학회지
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• 제34권6호
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• pp.513-518
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• 1999

A total of 113 Vibrio sp. strains were examined for plasmid content which were subjected to digestion with restriction enzymes. About the 55% Vibrio spp. have the plasmid more than one. Most of these plasmid various derivatives ranged from $2.4\;kb{\sim}23\;kb$, especially two strains of V. mimicus and one strain of V. furnissii carried one high-molecular weight plasmid (molecular weight ranging between $70\;kb{\sim}100\;kb$). Results of restriction analysis for plasmid of this three strains were by no means the rule. For detection of tdh and ctx gene, the virulence factor involved in the pathogenesis, we carried out the TDH and CT assay, PCR amplification, and hybridization. A total 11 strains were produced TDH, involved in 9 strains of V. parahaemolyticus and 1 strain of V. alginolyticus from clinical isolates and 1 strains of V. mimicus from environmental isolates. In the experiments of tdh gene detection, in all, 3 strains of V. parahaemolyticus from clinical isolates and 2 strains from environmental isolates could be successfully amplified in 400 bp by PCR. The PCR results were consistent with DNA hybridization tests. In the experiments of CT assay, in all, 3 strains of V. cholerae from clinical isolate and 1 strain of V. cholerae from environmental isolates were observed CT-producing. These CT-producing strains amplified in 302 bp by PCR for the detection of ctx gene. All CT-producing strains hybridized with digoxigenin-labeled DNA probe, while CT-negative strains did not hybridize. Also hybridization tests results for detection of ctx gene consistent with PCR.

• Journal of Microbiology and Biotechnology
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• 제22권7호
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• pp.889-893
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• 2012

Appressorium is a specialized infection structure of filamentous pathogenic fungi and plays an important role in establishing a pathogenic relationship with the host. The Egh16/Egh16H family members are involved in appressorium formation and pathogenesis in pathogenic filamentous fungi. In this study, a homolog of Egh16H, Magas1, was identified from an entomopathogenic fungus, Metarhizium acridum. The Magas1 protein shared a number of conserved motifs with other Egh16/Egh16H family members and specifically expressed during the appressorium development period. Magas1-EGFP fusion expression showed that Magas1 protein was not localized inside the cell. Deletion of the Magas1 gene had no impact on vegetative growth, conidiation and appressorium formation, but resulted in a decreased mortality of host insect when topically inoculated. However, the mortality was not significant between the Magas1 deletion mutant and wild-type treatment when the cuticle was bypassed by injecting conidia directly into the hemocoel. Our results suggested that Magas1 may influence virulence by affecting the penetration of the insects' cuticle.

• 대한수의학회지
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• 제39권2호
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• pp.327-337
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• 1999

The transposon TnphoA was used to generate avirulent mutants from a type A Pasteurella multocida. A suicide vector plasmid pRT733 carrying TnphoA, having the kanamycin resistant gene and harbored in Escherichia coli K-12 strain SM10(${\lambda}pir$), was mated with streptomycin resistant P. multocida P-1059 strain as recipient. This resulted in the generation of two TnphoA insertion mutants (transconjugants, tc95-a and tc95-b) which were resistant both to kanamycin ($Km^{R}$) and streptomycin ($Sm^{R}$), secreted alkaline phosphatase, and were avirulent to turkeys. Southern blot hybridization using two probes derived from internal fragments of TnphoA, confirmed the insertion of TnphoA into 12.9kb or 13.7kb DNA fragment from the EcoRV digested genomic fragments of transconjugants. The two transconjugants, tc95-a and tc95-b, were distinguishable from their parent strains by differences in ribotypes, and outer membrane protein profiles. TnphoA insertion in both transconjugants also resulted in constitutive expression of a 33Kd iron regulated outer membrane protein (IROMP). The gene encoding $Sm^{R}$ was also located within the same 12.9kb EcoRV genomic fragment from both transconjugants. Furthermore, our finding that the recipient P. multocida P-1059 $Sm^{R}$ strain and both transconjugants were avirulent to turkeys suggest that the either 12.9kb or 13.7kb genomic DNA contains the virulence gene and speculate that the presence of $Sm^{R}$ gene or TnphoA insertion may be responsible for regulating and inactivating the gene(s) encoding virulence in P. multocida.

• Journal of Veterinary Science
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• 제21권2호
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• pp.20.1-20.13
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• 2020

Actinobacillus pleuropneumoniae (APP) causes a form of porcine pleuropneumonia that leads to significant economic losses in the swine industry worldwide. The apxIBD gene is responsible for the secretion of the ApxI and ApxII toxins and the pnp gene is responsible for the adaptation of bacteria to cold temperature and a virulence factor. The apxIBD and pnp genes were deleted successfully from APP serotype 1 and 5 by transconjugation and sucrose counter-selection. The APP1ΔapxIBDΔpnp and APP5ΔapxIBDΔpnp mutants lost hemolytic activity and could not secrete ApxI and ApxII toxins outside the bacteria because both mutants lost the ApxI- and ApxII-secreting proteins by deletion of the apxIBD gene. Besides, the growth of these mutants was defective at low temperatures resulting from the deletion of pnp. The APP1ΔapxIBDΔpnp and APP5ΔapxIBDΔpnp mutants were significantly attenuated compared with wild-type ones. However, mice vaccinated intraperitoneally with APP5ΔapxIBDΔpnp did not provide any protection when challenged with a 10-times 50% lethal dose of virulent homologous (APP5) and heterologous (APP1) bacterial strains, while mice vaccinated with APP1ΔapxIBDΔpnp offered 75% protection against a homologous challenge. The ΔapxIBDΔpnp mutants were significantly attenuated and gave different protection rate against homologous virulent wild-type APP challenging.

• 생명과학회지
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• 제21권2호
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• pp.227-234
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• 2011

Pseudomonas syringae pv. tabaci는 숙주인 담배에 감염하여 들불병(wild fire)을 일으키는 식물 병원성 세균이다. 이 세균의 pathogenicity island (PAI)는 Type III secretion system 및 병원성 유전자들을 암호화하고 있으며, 병원성 조절에 있어 핵심적인 역할을 한다. 최근 식물 병원성 세균인 Ralstonia solanacearum에서 식물 세포 접촉을 매개로 하여 hrp gene cluster를 양성조절하는 PrhA (plant regulator of hrp) receptor가 발견되었다. 본 연구에서는 P. syringae에서 식물세포에 의해 hrp 유전자가 유도되는지 확인하기 위해, prhA 유사체를 동정하고 PrhA 결실돌연변이주(BL11)를 구축하였다. BL11은 숙주 감염 실험에서 병원성이 현저히 감소하였고, 식물 세포현탁액에서 hrpA 유전자의 발현수준이 hrp 유도배지에서 보다 3배 더 높게 나타났다. 이러한 결과들을 근거로 PrhA가 식물세포접촉에 의한 조절에 중요한 역할을 한다는 것을 확인하였으며, hrpA-gfp reporter fusion을 사용하여 이를 다시 검증하였다.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2005년도 제45회 학술심포지움 및 추계학술대회
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• pp.110-110
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• 2005