• 제목/요약/키워드: Toxin gene

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Phytotoxins of Pseudomonas syringae and PCR Primers for Detection of Phytotoxin-Producing Strains (Pseudomonas syringae의 식물독소와 독소 생산 균주의 검출을 위한 PCR Primer)

  • 정재성;한효심;고영진
    • Research in Plant Disease
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    • v.7 no.3
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    • pp.123-133
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    • 2001
  • Many pathovars of the species Pseudomonas syringae are known to produce different phytotoxins as secondary metabolites. Although phytotoxins generally enhance the virulence of P. syringae, they are not required for pathogenesis. Among the phytotoxins produced by P. syringae, lipodepsipeptides, coronatine, phaseolotoxin, and tabtoxin are the most well-known toxins which have been intensively studied for their structure, mode of action, biosynthesis, and regulation. A polymerase chain reaction (PCR) technique that amplifies a segment of the phytotoxin gene cluster using a primer set has been developed in recent years. This method offers the advantages of speed and sensitivity compared to the approaches based on physiological and biochemical methods. PCR detection of genes involved in the production of toxins could be exploited for early diagnosis of plant diseases caused by P. syringae pathovars.

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Identification of Novel Crystal Protein Gene from a Strain of Bacillus thuringiensis subsp. kenyae

  • Park, Jae-Young;Roh, Jong-Yul;Li, Ming-Shun;Shim, Hee-Jin;Boo, Kyung-Saeng;Je, Yeon-Ho
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.73-73
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    • 2003
  • Bacillus thuringiensis 2385-1, which showed toxicity to lepidopteran but not to dipteran was isolated from Korean soil sample and characterized. The H-serotype of 2385-1 was identical to that of serovar kenyae (H4a4c), and its crystal toxin was bipyramidal-shaped with a molecular weight of 130 kDa. However, the plasmid profile of 2385-1 was different from that of serovar kenyae. (omitted)

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Effect of Benzo(k)fluoroanthene and Genistein on CYP1A1 Gene Expression in Human Breast Cancer MCF-7 Cells. (사람 유방암 세포 MCF-7에서 Benzo(k)fluoroanthene과 genistein이 CYP1A1 유전자 발현에 미치는 영향)

  • Yang, So-Yeon;Min, Kyung-Nan;Shin, Yhun-Yhong
    • Environmental Mutagens and Carcinogens
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    • v.24 no.3
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    • pp.128-136
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    • 2004
  • CYP1A1 is known to be inducible by xenobiotic compouds such as polyciclic aromatic hydrocarbons(PAHs) and 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). These chemicals have been identified worldwide and can have a significant impact on the human health and well being of human and wildlife. Given these issues, the detection and quantification of these chemicals in biological, environmental and food samples is important. First, we investigated the effect of on CYP1A1 promoter activity, 7-ethoxyresorufin-O-deethylase(EROD) activity and CYP1A1 mRNA expression induced by benzo(k)fluoranthene(B(k)F) in MCF-7 cells. We found that B(k)F significantly up-regulates the level of CYP1A1 prompter activity, EROD and CYP1A1 mRNA. When cells were treated with genistein, it was not changed that EROD and CYP1A1 mRNA, compared to that of control. However, genistein inhibited the B(k)F-induced CYP1A1 promoter activity and mRNA level at high concentration. Furthermore, in this study, effects of HDAC(histone deacetvlase) inhibitors on human prostate cancer cells proliferation were examined. HC-toxin, SAHA and TSA inhibited cell proliferation in PC3 cells. A novel HDAC inhibitor, IN2001 also suppressed the growth of PC3 cells. And IN2001 and SAHA increased S phase and G2/M phase at 12 hrs treatment but cells were arrested G0/G1 phase at 45 hrs treatment. The HC-toxin treatment for 24 hrs and 48 hrs increased G0/G1 at low concentration ($0.1\mu\textrm{m}$) but increased G2/M at more than concentration of $1\mu\textrm{m}$. TSA increased G2/M phase. These findings height the possbility of developing HDAC inhibitors as potential anticancer therapeutic agents for the treatment of prostate cancer.

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Induction of a systemic IgG and secretory IgA responses in mice by peroral immunization with uropathogenic Escherichia coli adhesin protein coupled to cholera toxin A2B subunits

  • Lee, Yong-Hwa;Kim, Byung-Oh;Rhee, Dong-Kwon;Pyo, Suh-Kneung
    • Biomolecules & Therapeutics
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    • v.11 no.3
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    • pp.157-162
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    • 2003
  • The generation of secretory IgA antibodies(Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the ctxa2b gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimHIctxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and western blotting using antibodies to the maltose binding protein (MBP) or the cholera toxin subunit B (CTXB), plus the N-terminal amino acid sequence was analyzed. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/CTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of CTXB. This study also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked CTXA2B acts as a useful mucosal adjuvant, and that the adhesin/CTXA2B chimeric protein might be a potential antigen for oral immunization against UPEC.

Evaluation of Enterotoxigenic Bacteroides fragilis from Colonic Washings from Patients Undergoing Colonoscopy

  • Van, Ni;Ahlberg, Ned;Jung, Byung Chul;Lee, Min Ho;Ahn, Seung Ju;Lee, In-Soo;Kim, Yoon Suk;Rhee, Ki-Jong
    • Biomedical Science Letters
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    • v.18 no.4
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    • pp.362-368
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    • 2012
  • Enterotoxigenic Bacteroides fragilis (ETBF) is an intestinal commensal bacterium implicated as a risk factor for colon cancer. The key virulence factor is a secreted toxin called B. fragilis toxin (BFT). In this study we used an in vitro bioassay to examine the prevalence of ETBF in colonic washings from patients with colorectal polyps and normal control patients. We found that 9.3% of polyp patients and 10.9% of non-polyp patients harbored ETBF, respectively. A total of nine ETBF clinical isolates were isolated and confirmed to be positive for the BFT gene by PCR analysis and the ability to induce IL-8 secretion in the colonic epithelial cell line HT29/c1. Two of the ETBF clinical strains were characterized further in vitro and in vivo. We found that the two ETBF clinical isolates induced E-cadherin cleavage in HT29/c1 cells and promoted colonic inflammation in C57BL/6 mice. Our results indicate that the prevalence of ETBF in polyp patients were similar in non-polyp patients suggesting that ETBF carriage does not positively correlate to polyp incidence.

PCR technique for detection of toxigenic Pasteurella multocida in mixed bacterial cultures from pigs (Polymerase chain reaction을 이용한 독소생산성 Pasteurella multocida의 검출)

  • Chi, Yongzhe;Lee, Dong-seok;Han, Jeong-hee;Han, Kyung-soo;Hahn, Tae-wook
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.56-62
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    • 2000
  • Pasteurella multocida is kind of commensal bacteria in the upper respiratory tract of pigs. It is classified toxigenic and nontoxigenic strains based on the production of dermonecrotic toxin. Toxigenic strain is most associated with atrophic rhinitis which brings great economical loss in swine industry. However, toxigenic and nontoxigenic strains do not differ by diagnostic biochemical reaction or morphology. One of recently developed techniques, PCR detects the toxigenic P multocida. Amplification of an 846-nucleotide fragment of toxA gene was developed. The fragment amplified by PCR was detected in P multocida type D not type A. The PCR amplification was as sensitive as it could detect 1 pg of P multocida DNA. We compared the result of the PCR with the enzyme linked immunosorbent assay (ELISA) in a test for 40 swine nasal swabs. All of these isolates were toxin negative based on the ELISA while 2 isolates were detected in the PCR technique. in addition to accuracy, as required for rapid detection from contaminated nasal swabs, toxigenic P multocida was recovered efficiently from contaminated culture without inhibition of the PCR. The results show that the PCR detection of toxigenic P multocida directly form nasal swabs are feasible.

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Prevalence of virulence and cytolethal distending toxin (CDT) genes in thermophilic Campylobacter spp. from dogs and humans in Gyeongnam and Busan, Korea

  • Cho, Hyun-Ho;Kim, Sang-Hyun;Min, Wongi;Ku, Bok-Kyung;Kim, Yong-Hwan
    • Korean Journal of Veterinary Research
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    • v.54 no.1
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    • pp.39-48
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    • 2014
  • The prevalence of thermophilic Campylobacter (C.) spp. in stray, breeding, and household dogs was 25.2, 12.0, and 8.8%, respectively. C. jejuni and C. upsaliensis were the predominant Campylobacter spp. from household dogs. cdtA, cdtB, and cdtC were detected by PCR in all isolates. Despite the high cytolethal distending toxin (CDT) gene prevalence, only 26 (31%) C. jejuni strains and one (15.3%) C. coli strain showed evidence of CDT production in HEp-2 cell cytotoxicity assays. Virulence-associated genes detected in the C. jejuni and C. coli isolates were cadF, dnaJ, flaA, racR, ciaB, iamA, pldA, virB11, ceuE, and docC. cadF, dnaJ, flaA, and ceuE were found in all C. jejuni and C. coli isolates. When detecting Guillain-Barr$\acute{e}$ syndrome-associated genes (galE, cgtB, and wlaN), galE was identified in all isolates. However, cgtB and wlaN were more prevalent in C. jejuni isolates from humans than those from dogs. Adherence and invasion abilities of the C. jejuni and C. coli strains were tested in INT-407 cells. A considerable correlation (adjusted $R^2$= 0.678) existed between adherence and invasion activities of the Campylobacter spp. isolates.

Mosquitocidal Proteins from Escheriachia coli pSL 2-1 Clone and Bacillus sphaericus 1593 (Escheriachia coli pSL 2-1 클론과 Bacillus sphaericus 1593 균주가 생산한 모기치사 단백질)

  • Lee, Hong-Sup;Kim, Soo-Young;Lee, Hyung-Hoan
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.389-392
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    • 1988
  • A clone pSL 2-1, which is a recombinant plasmid believed to contain the mosquitocidal crystal-line protein gene of the Bacillus sphaericus 1593, was expressed in Escherichia coli JM83 and the product of the clone was purified and identified. The unsolubilized mosquitocidal crystal proteins from the B. sphaericus had formed 43, 58, 64, 100, 113, and 130 Kd bands in the SDS-polyacrylamide gel, but the NaOH-solublized proteins at pH 12 formed 2 protein bands of 43- and 64Kd in the gel because the larger protein (precursor) bands were cleaved. The products of the pSL 2-1 clone was purified by Sephadex G-200 and only the fractions having lethal activity to the 3rd in-star larvae of mosquito Culex pipiens were analyzed by the gel. The only single protein band of 42 Kd toxic to the larvae was formed. The major toxic protein being produced from the B. sphaericus 1593 and the pSL 2-1 clone was found to be the 42 Kd.

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Safety Assessment of Lactiplantibacillus (formerly Lactobacillus) plantarum Q180

  • Kwon, Yoo Jin;Chun, Byung Hee;Jung, Hye Su;Chu, Jaeryang;Joung, Hyunchae;Park, Sung Yurb;Kim, Byoung Kook;Jeon, Che Ok
    • Journal of Microbiology and Biotechnology
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    • v.31 no.10
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    • pp.1420-1429
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    • 2021
  • The safety of the probiotic strain Q180, which exerts postprandial lipid-lowering effects, was bioinformatically and phenotypically evaluated. The genome of strain Q180 was completely sequenced, and single circular chromosome of 3,197,263 bp without any plasmid was generated. Phylogenetic and related analyses using16S rRNA gene and whole-genome sequences revealed that strain Q180 is a member of Lactiplantibacillus (Lp., formerly Lactobacillus) plantarum. Antimicrobial resistance (AMR) genes were bioinformatically analyzed using all Lp. plantarum genomes available in GenBank, which showed that AMR genes are present differently depending on Lp. plantarum strains. Bioinformatic analysis demonstrated that some mobile genetic elements such as prophages and insertion sequences were identified in the genome of strain Q180, but because they did not contain harmful genes such as AMR genes and virulence factor (VF)- and toxin-related genes, it was suggested that there is no transferability of harmful genes. The minimum inhibition concentrations of seven tested antibiotics suggested by the European Food Safety Authority guidelines were slightly lower than or equal to the microbiological cut-off values for Lp. plantarum. Strain Q180 did not show hemolytic and gelatinase activities and biogenic amine-producing ability. Taken together, this study demonstrated the safety of strain Q180 in terms of absence of AMR genes and VF- and toxin-related genes as a probiotic strain.

Identification of Toxin Gene and Antibiotic Resistance of Staphylococcus Aureus Isolated from Agricultural Product Cultivation Environments (농산물 생산 환경에서 분리된 Staphylococcus aureus의 항생제 내성 및 독소 유전자 확인)

  • Park, Su-Hee;Kim, Jeong-Sook;Kim, Kyeong-Yeol;Chung, Duck-Hwa;Shim, Won-Bo
    • Journal of Environmental Health Sciences
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    • v.39 no.5
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    • pp.465-473
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    • 2013
  • Objectives: This study was undertaken to analyze Staphylococcus aureus from cultivation environments for agricultural products and to confirm antibiotic resistance and enterotoxin genes for the isolated S. aureus. Methods: A total of 648 samples were collected from apple, peach, ginseng and balloon flower farms. S. aureus was isolated from soil, agricultural water, personal hygiene elements (hands, gloves and clothes) and work utensils (boxes). Results: S. aureus was detected in a total of 25 samples and 72 strains were isolated. The resistance rate of the isolated S. aureus strains was confirmed at 33.3%, with 24 resistant strains among the total of 72. Fourteen different patterns types were found, and three pattern types (NV, OX, VA) were confirmed most frequently. As result of the detection of enterotoxin gene type, four gene types (sea: 1, sed: 4, seg: all isolated S. aureus, sei: all isolated S. aureus) were analyzed among a total of nine types. Conclusions: This study demonstrates that personal hygiene techniques should be properly managed, such as washing and sterilization before or after work, because agricultural contamination by S. aureus frequently developed through improper management.