• Title/Summary/Keyword: Virulence gene

Search Result 312, Processing Time 0.026 seconds

Cloning, Sequencing and Expression of apxIA, IIA, IIIA of Actinobacillus pleuropneumoniae Isolated in Korea (국내 분리 흉막폐렴균의 apxIA, IIA, IIIA 유전자 Cloning, 염기서열 분석 및 단백질 발현)

  • Shin, Sung-jae;Cho, Young-wook;Yoo, Han-sang
    • Korean Journal of Veterinary Research
    • /
    • v.43 no.2
    • /
    • pp.247-253
    • /
    • 2003
  • Actinobacillus pleuropneumoniae causes a highly contagious pleuropneumoniae in swine. The bacterium produces several virulence factors such as exotoxin, LPS, capsular polysaccharide, etc. Among them, the exotoxin, called Apx, has been focused as the major virulence factor, and the toxin consists of 4 gene cluster. apx CABD. apxA is the structural gene of toxin and has four different types, I, II, III, and IV. As the first step of development of a new subunit vaccine, the three different types of apxA gene were amplified from A. pleuropneumoniae isolated from Korea by PCR with primer designed based on the N- and C-terminal of the toxin. The sizes of apxIA, IIA and IIIA were 3,073, 2,971 and 3,159bps, respectively. The comparison of whole DNA sequences of apxIA, IIA and IIIA genes with those of the reference strain demonstrated 98%, 99% and 98% homology, respectively. In addition, the phylogenetic analysis was performed based on the amino acid sequences compared with 12 different RTX toxin family using the neighbor-joining method. ApxA proteins of Korean isolates were identical with reference strains in this study. All ApxA proteins were expressed in E. coli with pQE expression vector and identified using Western blot with polyclonal antibodies against culture supernatants of A. pleuropneumoniae serotype 2 or 5. The sizes of each expressed ApxA protein were about 120, 110, 125 kDa (M.W.), respectively. The results obtained in this study could be used for the future study to develop a new vaccine to porcine pleuropneumoniae.

Race Classification of the Bacterial Blight Pathogen, Xanthomonas oryzae pv. oryzae, by Rice NILs with Single Resistance Genes (벼 흰잎마름병 저항성 유전자를 갖고 있는 준동질 계통을 이용한 벼 흰잎마름병균의 레이스 분류)

  • Choi, Jae-Eul;Kim, Bo-Ra;Han, Jin-Soo;Kang, Hee-Kyoung;Hur, Seung-Gi
    • Research in Plant Disease
    • /
    • v.14 no.3
    • /
    • pp.165-170
    • /
    • 2008
  • One hundred and three isolates of Xanthomonas oryzae pv. oryzae in Korea were evaluated for their virulence on four near-isogenic lines (NILs) containing a single resistance gene, and Korean differential varieties. The resistant gene backgrounds of Cheongcheongbyeo, Pungsanbyeo, Hangangchalbyeo, Milyang42 were not completely understood and they were not suited for the classification of X. oryzae pv. oryzae. Four NILs, IRBB101, IRBB103, IRBB105, and IRBB107 were difference for characterizing races of X. oryzae pv. oryzae because they have a single resistance gene. These NILs may be useful differential set in examining pathogenic races of X. oryzae pv. oryzae in Korea. Based on the virulence of 103 isolates to new differential varieties, they were classified into 3 races.

Charaterization of an Escherichia coli O157:H7 Strain Producing Verotoxin 2Isolated from a Patient in Korea

  • Park, Wan;Sohn, Chang-Kyu;Wan Huh;Kim, Byung-Chun
    • Journal of Microbiology
    • /
    • v.38 no.2
    • /
    • pp.93-98
    • /
    • 2000
  • Nine hundred patients diagnosed with diarrhea or hemorrhagic uremic syndrome in the Kyungpook Province, Korea, were examined from November 1998 to February 2000. One patient in Kumi appeared to possess the Escherichia coli O157:H7 strain, which is very important in clinical decision making and public health action. The isolated strain, an E. coli O157:H7 KM, contained a 60 MDa plasmid and typical virulence genes including the verotoxin 2 gene, ehxA gene (encoding enterohemorrhagic hemolysin), and eae (encoding attaching and effacing protein-intimin) gene. This strain produced only verotoxin 2. Pulsed field gel electrophoretic analysis showed that the genomic organization of the E. coli O157:H7 KM strain may differ greatly from those of representative strains previously reported in the United States and Japan.

  • PDF

Detection of Vibrio vulnificus by Real-Time PCR targeted to rpoS gene (rpoS 유전자를 대상으로 하는 Real-Time PCR에 의한 Vibrio vulnificus 검출)

  • Kim, Dong-Gyun;Ahn, Sun-Hee;Bae, Ju-Yoon;Kong, In-Soo
    • Journal of Marine Bioscience and Biotechnology
    • /
    • v.2 no.4
    • /
    • pp.263-266
    • /
    • 2007
  • Vibrio vulnificusis a causative agent of serious diseases in humans resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium using the rpoS gene in pure cultures and in infected oyster tissues.

  • PDF

INHIBITION OF GLUCAN SYNTHESIS RELATED GENE EXPRESSION OF STREPTOCOCCUS MUTANS BY XYLITOL TREATMENT (자일리톨 섭취에 따른 Streptococcus mutans의 글루칸 생성관련 유전자 발현 억제효과)

  • Kim, Ji-Hye;Lee, Young-Eun;Ahn, Sang-Hun;Choi, Youn-Hee;Nam, Soon-Heyun;Song, Keun-Bae
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.36 no.4
    • /
    • pp.531-538
    • /
    • 2009
  • Xylitol has the ability to reduce the adherence of Streptococcus mutans(S. mutans), which can make it easier to remove plaque, decrease acid production and inhibit dental caries. There are few reports on the effects of xylitol on the expression of the virulence related genes in S. mutans. This study examined the inhibitory effect of chewing gum containing xylitol on glucan synthesis related gene expression of S. mutans. Participants were voluntarily recruited for a women's oral health prevention program, classified into two groups(a control and a xylitol group), and then followed for 2 years. Twenty salivary samples were randomly selected from each group. Colony count and real-time reverse transcription polymerase chain reaction were used to analyze the characteristics of S. mutans. The following results were obtained: The S. mutans counts decreased steadily in the xylitol group over the study period(p<0.05). The expression of the virulence related genes (gtfB, gtfC and gtfD) was significantly lower in the xylitol group than in the control groups (p<0.05). In conclusion, these results suggest that chewing xylitol gum for a long period of time may reduce the expression of the genes associated with S. mutans virulence, which can result in a decrease growth of S. mutans colonies as a result.

  • PDF

Role of Salmonella Typhimurium SlyA in Regulating the Expression of VirulenceFactors Related to Survival in Macrophages (대식세포 내 생존과 관련된 독성인자 발현 조절에서의 Salmonella Typhimurium SlyA의 역할)

  • Yeo bin Kim;Jeong-eun Baek;Yeon Ha Kim;Young Hee Kim;Ah Young Yoo;Ho Young Kang
    • Journal of Life Science
    • /
    • v.34 no.2
    • /
    • pp.79-85
    • /
    • 2024
  • SlyA is known as a transcriptional regulator that regulates the expression of hemolysin (HlyE) in E. coli, a member of the Enterobacteriaceae family such as Salmonella. However, Salmonella has the slyA gene but lacks the hlyE gene. Then, because we were curious about the role of SlyA in Salmonella, we constructed and explored a mutant strain with a deletion of the slyA gene. S. Typhimurium CK295 (ΔslyA) was constructed using an allelic exchange approach. In a comparative analysis between the wild-type and the CK295 strain, no significant differences were observed in growth characteristics, motility, total protein analyses, and secreted protein analyses. However, the CK295 strain exhibited slightly reduced biofilm formation compared to the wild-type. Interestingly, as a result of comparing the survival ability in macrophages, the mutant strain showed a 60% decrease in survival ability compared to the wild-type. To evaluate toxicity in mice, mortality was measured after oral administration to 6-week-old BALB/c mice. As a result, the LD50 value of the CK295 (ΔslyA) was more than 100 times higher than that of wild-type S. Typhimurium 𝜒3339 in BALB/c. In conclusion, SlyA is presumed to regulate the expression of genes encoding virulence factors involved in the in vivo survival of Salmonella.

Genomic Insights into the Rice Blast Fungus through Estimation of Gene Emergence Time in Phylogenetic Context

  • Choi, Jaeyoung;Lee, Jong-Joon;Jeon, Junhyun
    • Mycobiology
    • /
    • v.46 no.4
    • /
    • pp.361-369
    • /
    • 2018
  • The rice blast fungus, Magnaporthe oryzae, is an important pathogen of rice plants. It is well known that genes encoded in the genome have different evolutionary histories that are related to their functions. Phylostratigraphy is a method that correlates the evolutionary origin of genes with evolutionary transitions. Here we applied phylostratigraphy to partition total gene content of M. oryzae into distinct classes (phylostrata), which we designated PS1 to PS7, based on estimation of their emergence time. Genes in individual phylostrata did not show significant biases in their global distribution among seven chromosomes, but at the local level, clustering of genes belonging to the same phylostratum was observed. Our phylostrata-wide analysis of genes revealed that genes in the same phylostratum tend to be similar in many physical and functional characteristics such as gene length and structure, GC contents, codon adaptation index, and level of transcription, which correlates with biological functions in evolutionary context. We also found that a significant proportion of genes in the genome are orphans, for which no orthologs can be detected in the database. Among them, we narrowed down to seven orphan genes having transcriptional and translational evidences, and showed that one of them is implicated in asexual reproduction and virulence, suggesting ongoing evolution in this fungus through lineage-specific genes. Our results provide genomic basis for linking functions of pathogenicity factors and gene emergence time.

Investigation on antimicrobial resistance genes of Salmonella spp. isolated from pigs and cattle (소와 돼지유래 살모넬라속균의 약계내성유전자의 특성에 관한 연구)

  • Lee, Woo-Won;Jung, Byeong-Yeal;Lee, Gang-Rok;Lee, Dong-Soo;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
    • /
    • v.32 no.3
    • /
    • pp.227-239
    • /
    • 2009
  • At the present study, it was aimed to detect virulence genes and antimicrobial resistance genes among 102 strains of 12 Salmonella serotypes isolated from pigs and cattle. In polymerase chain reaction (PCR), invA was detected from all strains of Salmonella spp., spvC was detected from Salmonella enterica serotype Enteritidis (S. Enteritidis) (100%), S. Bradenburg (75%), and S. Typhimurium (20.4%). Drug resistance related genes of 12 types were detected from all strains. TEM ($bla_{TEM}$) gene was detected from 51 (92.7%) of 55 $\beta$-lactams (54 ampicillin or 1 amoxicillin) resistance strains. 55 (100%) of 55 chloramphenicol resistance strains, 3 (100%) of 3 gentamicin resistance strains and 5 (100%) of 5 kanamycin resistance strains did contain cml, aadB, and aphA1-Iab, respectively. strB (89.9%), strA (88.4%), aadA2 (84.1%) and aadA1 (72.5%) were detected from 69 streptomycin resistance strains. sulII and dhfrXII were detected from 49 (100%) of 49 sulfamethoxazole/trimethoprim resistance strains, but sulI was not detected. tetA (97.9%) and tetB (21.6%) were detected from 97 tetracycline resistance strains. int gene was detected from 58 (56.9%) of 102 strains. 54 S. Typhimurium of 102 Salmonella spp. were attempted to detect drug resistance genes. TEM was detected from 44 (95.7%) of 46 $\beta$-lactams (45 ampicillin or 1 amoxicillin) resistance strains. cmlA was detected from 51 (100%) of 51 chloramphenicol resistance strains. aadA2 (100%), strA (100%), strB (100%), and aadA1 (79.6%) were detected from 54 streptomycin resistance strains. sulII (100%) and dhfrXII (100%) were detected from 49 sulfamethoxazole/trimethoprim resistance strains. tetA was detected from 54 (100%) of 54 tetracycline resistance strains. int gene was detected from 54 (100%) of 54 strains. The major drug resistance pattern and resistance gene profile were ampicillin, chloramphenicol, streptomycin, sulfamethoxazole/trimethoprim and tetracycline (ACSSuT) and TEM, cmlA, aadA1, aadA2, strA, strB, sulII, dhfrXII, tetA and int, respectively.

Transcriptional Responses of Respiratory Epithelial Cells to Nontypable H. influenzae Infection: Identification of Differentially Regulated Genes by Microarray Analysis of Human cDNA

  • Lee, Na-Gyong
    • Proceedings of the PSK Conference
    • /
    • 2002.10a
    • /
    • pp.151-152
    • /
    • 2002
  • Bacterial infection is a very complex process in which both pathogenic microorganisms and host cells play crucial roles, and it is the outcome of interactions between the two participants. To elucidate the bacterial pathogenesis mechanisms, therefore, it is essential to understand the cellular and systemic responses of the host as well as the virulence factors of the pathogen. Infection of a host by pathogenic bacteria causes drastic changes in the physiology of host cells, leading to activation of a program of various gene expression. (omitted)

  • PDF

AHL inhibition of Beckerelide and Fimbrolide

  • Kim, Yeon-Hee;Lee, Jae-Gun;Park, Sung-Hoon;Kim, Jung-Sun
    • Proceedings of the PSK Conference
    • /
    • 2003.10b
    • /
    • pp.174.2-174.2
    • /
    • 2003
  • Quorum sensing, a gene expression in response to population density, is regulated by chemical signals, most of which are acylated homoserine lactones (AHLs). The AHL derivatives have been reported to regulate bioluminescence, virulence factors and / or swarming motility in bacteria. It is hypothesized that higher organisms may have evolved specific means to interfere with bacterial communication as exemplified in the AHL-antagonistic activity of halogenated furanones isolated from the Australian macroalga Delisea pulchra. (omitted)

  • PDF