• 제목/요약/키워드: toxA gene

검색결과 28건 처리시간 0.018초

Development of a toxA Gene Knock-out Mutant of Pasteurella multocida and Evaluation of its Protective Effects

  • Kim Tae-Jung;Lee Jae-Il;Lee Bong-Joo
    • Journal of Microbiology
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    • 제44권3호
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    • pp.320-326
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    • 2006
  • Pasteurella multocida is an important veterinary and opportunistic human pathogen. In particular, strains of P. multocida serogroup D cause progressive atrophic rhinitis, and produce a potent, intracellular, mitogenic toxin known as P. multocida toxin (PMT), which is encoded by the toxA gene. To further investigate the toxigenic and pathogenic effects of PMT, a toxA-deleted mutant was developed by homologous gene recombination. When administrated to mice, the toxigenicity of the toxA mutant P. multocida was drastically reduced, suggesting that the PMT constributes the major part of the toxigenicity of P, multocida. Similar results were obtained in a subsequent experiment, while high mortalities were observed when toxA(+) P. multocida bacterial culture or culture Iysate were administrated. Mice immunized with toxA(-) P. multocida were not protected (none survived) following challenge with toxA(+) P. multocida or bacterial culture Iysate (toxin). These results suggest that the toxigenicity of P. multocida is mainly derived from PMT.

분자생물학적 방법에 의한 남조류의 독성 생성능의 확인 (Detection of Toxigenicity of Cyanobacteria by Molecular Method)

  • 이경락;정원화;김종민;김한순
    • 생태와환경
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    • 제40권1호
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    • pp.149-154
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    • 2007
  • 남조류의 독성 잠재력을 확인하기 위해 microcystins 합성 유전자인 mcyB에 특이적으로 작용하는 TOX2P/TOX2M primer쌍을 이용한 whole-cell PCR을 실시하였다. 환경시료를 대상으로 한 실험 결과에서 TOX2P/TOX2M primer쌍은 약 1,000 $cells{\cdot}mL^{-1}$의 낮은 밀도에서 효과적으로 mcyB 유전자의 증폭에 성공하였으며, mcyB유전자를 지닌 종들은 모두 ELISA분석에 의해 microcystins의 생성이 확인되었다. 따라서, TOX2P/TOX2M primer쌍은 국내의 수체에서 독성 남조류의 신속하고 효과적인 검출을 위한 유용한 probe로 판단되었다.

돼지 폐렴병소로부터 분리한 Pasteurella multocida의 capsular serogroup과 toxA gene의 분포 (Capsular serogroups and toxA gene of Pasteurella multocida isolated from Pneumonic Lung Lesions of Swine)

  • 손준형;최성균;조길재
    • 한국임상수의학회지
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    • 제26권5호
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    • pp.457-462
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    • 2009
  • The present study was conducted to investigate the capsular serogroups and distribution of toxA gene of Pasteurella (P.) multocida isolated from pneumonic lung lesions of swine in Korea. A total number of 91 (36.3%) P. multocida isolated from 251 lung lesions. P. multocida isolates were typed for capsular serogroup and toxA gene by polymerase chain reaction. Of the 91 strains, serogroup A and D were 69 strains (75.8%) and 22 strains (24.2%), respectively. Sixty one strains (67.0%) out of 91 strains were detected as toxA gene, and 47 strains (77.0%) and 14 strains (23.0%) belongs to serogroup A and D, respectively.

Galleria mellonella 6-Tox Gene, Putative Immune Related Molecule in Lepidoptera

  • Lee, Joon-Ha;Park, Seung-Mi;Chae, Kwon-Seok;Lee, In-Hee
    • International Journal of Industrial Entomology and Biomaterials
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    • 제21권1호
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    • pp.127-132
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    • 2010
  • We have characterized full-length cDNA encoding Gall-6-tox protein, which was cloned from the fat body of the immunized Galleria mellonella larvae. The cloned cDNA of Gall-6-tox consists of 1301 nucleotides and contained an open reading frame of 891 nucleotides corresponding to a protein of 296 residues that includes a putative 16-residue signal sequence and a 280-residue mature peptide with a calculated mass of 30,707.73 Da. The deduced mature peptide contains conserved tandem repeats of six cysteine-stabilized alpha beta ($Cs{\alpha}{\beta}$) motifs, which was detected in scorpion toxins and insect defensins. In the sequence homology search, mature Gall-6-tox showed 34% and 28% amino acid sequence homology with Bomb-6-tox from Bombyx mori and Spod-11-tox from Spodoptera frugiperda, respectively. Gall-6-tox orthologs were only found in Lepidopteran species, indicating that this new immune-related gene family is specific to this insect order. RT-PCR analysis revealed that Gall-6-tox was expressed primarily in the larval fat bodies, hemocytes, and midgut against invading bacteria into hemocoel. Moreover, the expression time course of Gall-6-tox was examined up to 24 h in the fat bodies and midgut after injection of E. coli. Altogether, these results suggest that Gall-6-tox is derived from defensins and Gall-6-tox may play a critical role in Lepidoptera immune system.

돼지 폐렴병소로부터 분리한 Pasteurella multocida에 관한 연구 : 항균제 감수성, plasmid profile 및 toxA 유전자 분포 (Characteristics of Pasteurella multocia isolated from pneumonic lung lesions of swine ; antimicrobial susceptibility, plasmid profile and distribution of toxA)

  • 신나리;박주연;박용호;유한상
    • 대한수의학회지
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    • 제39권6호
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    • pp.1091-1098
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    • 1999
  • Antimicrobial susceptibility, plasmid profiles and distribution of toxA gene were investigated in Pasteurella multocida isolated from pneumonic lung lesions of swine. The bacteria were highly susceptible to norfloxacin, cabenicillin, enrofloxacin and chloramphenicol, but resistant to colistin, sulfamethoxawle/trimethoprime, bacitracin, streptomycin. Sixty percentage of the isolates was resistant more than 2 drugs used in this experiment and 21 strains (23.6%) were resistant more than 5 drugs. This phenomenon meant that they had highly multi-drugs resistance. In the analysis of plasmid profiles, nineteen strains (47.5%) of 40 P multocida isolates harbored plasmids, ranging from 53.3kb to 2.49kb in size and the plasmid profiles could be classified into 5 groups. However, there was no relationship between the size and the profile of plasmid and the resistance pattern of antimicrobial agents. Thirty strains of 39 P multocida isolates (77%) investigated by PCR harbored toxA gene. This result suggested involvement of the ToxA protein expressed from the gene in pneumonic pasteurellosis of swine.

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H-NS Silences Gene Expression of LeuO, the Master Regulator of the Cyclic(Phe-Pro)-dependent Signal Pathway, in Vibrio vulnificus

  • Park, Na-Young;Lee, Keun-Woo;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • 제30권6호
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    • pp.830-838
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    • 2020
  • The histone-like nucleoid structuring protein (H-NS) is an abundant global regulator of environmentally controlled gene expression. Herein, we demonstrate that H-NS represses the expression of LeuO, the master regulator of the cyclic(Phe-Pro)-dependent signaling pathway, by directly binding to the upstream region of the gene. H-NS binds to a long stretched region (more than 160-bp long), which overlaps with binding sites for ToxR and LeuO. A high quantity of H-NS outcompetes ToxR for binding to the cis-acting element of leuO. However, our footprinting analyses suggests that the binding of H-NS is relatively weaker than LeuO or ToxR at the same molarity. Considering that the DNA nucleotide sequences of the upstream regions of leuO genes are highly conserved among various Vibrio, such patterns as those found in V. vulnificus would be a common feature in the regulation of leuO gene expression in Vibrionaceae. Taken together, these results suggest that, in species belonging to Vibrionaceae, H-NS regulates the expression of leuO as a basal stopper when cFP-ToxR mediated signaling is absent.

Molecular Cloning of Two cDNAs Encoding an Insecticidal Toxin from the Spider, Araneus ventricosus, and Construction of a Recombinant Baculovirus Expressing a Spider Toxin

  • Chung, Eun-Hwa;Lee, Kwang-Sik;Han, Ji-Hee;Je, Yeon-Ho;Chang, Jin-Hee;Roh, Jong-Yul
    • International Journal of Industrial Entomology and Biomaterials
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    • 제4권1호
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    • pp.43-49
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    • 2002
  • We have cloned cDNAs encoding toxin from the spider, Araneus ventricosus, and constructed a recombinant baculovirus expressing the insecticidal toxin. The cDNAs encoding toxin were cloned from the cDNA library of A. ventricosus. Sequence analysis of the cDNAs encoding the toxin of A. ventricosus revealed that the 240 bp cDNA for AvTox-1 and 192 bp cDNA for AvTox-2 have an open reading frame of 80 and 64 amino acid residues, respectively. The deduced protein sequence of the toxin genes of AvTox-1 and AvTox-2 was aligned to that of the snack Anemonia sulcata and scorpion Centruroides limpidus limpidus, respectively. Northern blot analysis indicated that AvTox-2 toxin gene showed a fat body-spe-cific expression pattern at the transcriptional level. Furthermore, we have explored the possibility of improving baculovirus by incorporating the A. vontricosus toxin gene into Bombyx mori nuclear polyhedrosis virus genome under the control of polyhedrin promoter, The AvTox-2 toxin gene was expressed as approximately 5.8 kDa band in the recombinant baculovirus-injected silkworm larvae. Bioassays with the recombinant virus expressing AvTox-2 on 5th instar silkworm larvae demonstrated a decrease in the time to kill $(LT_{50} days)$ compared to wild-type BmNPV-Kl $(LT_{50} 6.72 days)$ in the injection of 10 viruses. These results indicate that A. ventricosus toxin is a novel member of the spider toxin family, suggesting that the toxin gene can be used in recombinant baculoviruses to reduce insect feeding damage and increase the speed of insect kill.

Virulence Phenotyping and Molecular Characterization of a New Virulence Type of Pyrenophora tritici-repentis the Causal Agent of Tan Spot

  • Benslimane, Hamida
    • The Plant Pathology Journal
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    • 제34권2호
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    • pp.139-142
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    • 2018
  • Pyrenophoratritici-repentis is the causal agent of tan spot. According to their ability to produce necrosis and/or chlorosis on a set of four differential bread wheats, the isolates of this fungus are currently grouped into eight races. When durum wheat genotypes were added to the differential set, a new virulence type was identified in Algeria. The isolates showing this virulence pattern are unable to attack bread wheat while they cause necrosis in durum genotypes. In this work, characterization of those isolates was based on pathological and molecular aspects. This included inoculation of bread and durum wheat, and virulence gene analysis using PCR and sequencing. The results showed that all isolates caused a resistance on all bread wheats of the differential set, while they produced necrosis in durum. ToxA and ToxB genes were amplified in all isolates, whereas toxb was absent. Sequence analysis for both genes showed no differences with those found in the two functional genes. The presence of two genes, ToxA and ToxB, despite the absence of symptoms usually caused by their products, suggests the existence of a new homologous for these two genes yet unknown. The presence of ToxA in the isolate unable to produce necrosis in Glenlea is reported for the first time.

독소 생성 Microcystis 검출을 위한 PCR primer의 평가 (Primer Evaluation for the Detection of Toxigenic Microcystis by PCR)

  • 이현경;김준호;유순애;안태석;김치경;이동훈
    • 미생물학회지
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    • 제39권3호
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    • pp.166-174
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    • 2003
  • Microcystin은 부영양화된 하천과 호수에서 cyanobacteria에 의해 생성되며 인간과 야생 동물에게 독성 물질로 작용하여 심각한 환경문제를 일으킨다. Microcystin은 mcy 유전자에 의해 암호화되는 microcystin synthetase로 알려진 multi-functional enzyme complex에 의해 리보좀의 관여 업이 합성된다. 따라서 mcy유전자의 PCR 증폭을 통해 독소를 생성하는 Microcystis를 효과적으로 검출할 수 있다. 본 연구에서는 microcystin을 생성하는 균주를 구별하기 위해 설계된 7종의 primer쌍의 유용성을 검증하기 위하여, 17주의 cyanobacteria와 국내 호수 시료에서 추출된 핵산을 대상으로 PCR 증폭을 하였다. TOX4E-TOX4R, FAA-RAA, FP-RP primer쌍에 의해서는 독소를 생성하는 Microcystis 균주 중 일부가 검출되지 않았다. NSZW2-NSZW1 primer쌍을 사용한 PCR의 경우 microcystin을 생성하는 국내 균주에서 예상하지 못한 크기의 산물이 관찰되었다. TOX1P-TOX1F primer쌍으로 PCR을 한 결과, 증폭된 산물을 관찰할 수 없었다. MSF-MSR과 TOX2P-TOX2F primer쌍만이 독소를 생성하는 11주의 Microcystis로부터 mcy유전자의 증폭을 성공하였다. 20개의 국내호수 시료에 각 Primer쌍에 의한 증폭여부를 확인한 결과, TOX2P-TOX2F primer쌍을 사용한 경우에만 모든 호수 시료에서 증폭된 산물을 관찰할 수 있었다. 본 연구 결과 TOX2P-TOX2F primer쌍이 국내 환경에서 독소를 생산하는 Microcystis를 검출하는데 가장 우수한 primer임을 확인할 수 있었다. 또한 Microcystis aenginosa NIER10010의 mcy 유전자의 염기서열 분석을 통해 국내 분리 균주의 유전적 다양성을 확인할 수 있었다.X> 을 일부 함유하고 있기 때문인 것으로 여겨진다. 궁극적으로 이 연구결과는 벤토나이트의 품위 산정에는 XRD 정량분석법이 적용되는 것이 합리적이고 CEC와 관련된 품질 특성은 전적으로 ‘Total CEC’ 개념에 의거하여 평가되어야 한다는 것을 시사한다.세균은 부유세균과는 다른 다양성을 이루고, 다른 천이과정을 거치는 것으로 확인되었다.로 interlayer된 것을 보여준다. 이와 같은 결과는 백운모, 녹니석 및 흑운모와 같은 변성 광물들이 비평형 광물 반응으로 만들어 졌으며 그 결과 불균질한 광물로 되었다는 것을 암시한다. led to the highest durability of all tested here. The reason of the improvement is due to thin MgF$_2$, which can prevent the $Mg_2$Ni electrode from forming Mg(OH)$_2$layer that is the main cause of degradation.platin에 의한 직접적 폐 독성은 발견되지 않았다이 낮았으나 통계학적 의의는 없었다[10.0%(4/40) : 8.2%(20/244), p>0.05]. 결론: 비디오흉강경술에서 재발을 낮추기 위해 수술시 폐야 전체를 관찰하여 존재하는 폐기포를 놓치지 않는 것이 중요하며, 폐기포를 확인하지 못한 경우와 이차성 자연기흉에 대해서는 흉막유착술에 더 세심한 주의가 필요하다는 것을 확인하였다. 비디오흉강경수술은 통증이 적고, 입원기간이 짧고, 사회로의 복귀가 빠르며, 고위험군에 적용할 수 있고, 무엇보다도 미용상의 이점이 크다는 면에서 자연기흉에 대해 유용한 치료방법임에는 틀림이 없으나 개흉술에 비해 재발율이 높고 비용이 비싸다는 문제가 제기되고 있는 만큼 더 세심한 주의와 장기 추적관찰이 필요하리라 사료된다.전 도부타민 심초음파는 관상동맥우회로술 후 동면심근의

Variations in the Antivirulence Effects of Fatty Acids and Virstatin against Vibrio cholerae Strains

  • Donghyun Lee;Jayun Joo;Hunseok Choi;Seonghyeon Son;Jonghyun Bae;Dong Wook Kim;Eun Jin Kim
    • Journal of Microbiology and Biotechnology
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    • 제34권9호
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    • pp.1757-1768
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    • 2024
  • The expression of two major virulence factors of Vibrio cholerae, cholera toxin (CT) and toxin co-regulated pilus (TCP), is induced by environmental stimuli through a cascade of interactions among regulatory proteins known as the ToxR regulon when the bacteria reach the human small intestine. ToxT is produced via the ToxR regulon and acts as the direct transcriptional activator of CT (ctxAB), TCP (tcp gene cluster), and other virulence genes. Unsaturated fatty acids (UFAs) and several small-molecule inhibitors of ToxT have been developed as antivirulence agents against V. cholerae. This study reports the inhibitory effects of fatty acids and virstatin (a small-molecule inhibitor of ToxT) on the transcriptional activation functions of ToxT in isogenic derivatives of V. cholerae strains containing various toxT alleles. The fatty acids and virstatin had discrete effects depending on the ToxT allele (different by 2 amino acids), V. cholerae strain, and culture conditions, indicating that V. cholerae strains could overcome the effects of UFAs and small-molecule inhibitors by acquiring point mutations in toxT. Our results suggest that small-molecule inhibitors should be examined thoroughly against various V. cholerae strains and toxT alleles during development.