• Title/Summary/Keyword: Bacillus Anthracis

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Curing Both Virulent Mega-Plasmids from Bacillus anthracis Wild-Type Strain A16 Simultaneously Using Plasmid Incompatibility

  • Wang, Dongshu;Gao, Zhiqi;Wang, Huagui;Feng, Erling;Zhu, Li;Liu, Xiankai;Wang, Hengliang
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1614-1620
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    • 2015
  • Plasmid-cured derivative strains of Bacillus anthracis are frequently used in laboratory studies. Plasmid incompatibility, which does not increase the risk of chromosomal mutation, is a useful method for plasmid curing. However, in bacteria containing multiple plasmids, it often requires the sequential introduction of multiple, specific incompatibility plasmids. This lengthy process renders the traditional plasmid incompatibility method inefficient and mutation-prone. In this study, we successfully cured plasmids pXO1 and pXO2 from B. anthracis A16 simultaneously using only one recombinant incompatible plasmid, pKORT, to obtain a plasmid-free strain, designated A16DD. This method may also be useful for the simultaneous, one-step curing of multiple plasmids from other bacteria, including Bacillus thuringiensis and Yersinia pestis.

Whole Transcriptomic Analysis of Bacillus anthracis during Hydrogen Peroxide Decontamination (과산화수소 제독 과정에서의 탄저균 전사체 분석)

  • Kim, Sang Hoon;Kim, Se Kye;Jung, Kyoung Hwa;Yoon, Sung Nyo;Kim, Yun Ki;Kim, Min Cheol;Ryu, Sam Gon;Lee, Hae Wan;Chai, Young Gyu
    • Journal of the Korea Institute of Military Science and Technology
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    • v.18 no.4
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    • pp.478-483
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    • 2015
  • Decontamination of biological agents utilizes hydrogen peroxide($H_2O_2$) for its effectiveness and safeness. Bacillus anthracis is a major target for $H_2O_2$ decontamination. To assess the effect of $H_2O_2$ on B. anthracis and identify biomarkers for decontamination, whole transcriptomic profiling of $H_2O_2$-treated B. anthracis was performed. Here we identified deregulation in stress response genes, transcription factors and cellular homeostasis genes. We also found that expression of antisense RNAs increased in B. anthracis during decontamination. We postulate that B. anthracis prioritizes survival and adaptation in response to $H_2O_2$ treatment by changing its gene expression pattern.

Expression of Acetohydroxyacid Synthase from Bacillus anthracis and Its Potent Inhibitors

  • Choi, Kyoung-Jae;Pham, Chien Ngoc;Jung, Hoe-Il;Han, Sung-Hwan;Choi, Jung-Do;Kim, Jin-Heung;Yoon, Moon-Young
    • Bulletin of the Korean Chemical Society
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    • v.28 no.7
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    • pp.1109-1113
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    • 2007
  • Acetohydroxyacid synthase (AHAS, EC 2. 2. 1. 6) is the enzyme that catalyses the first step in the common pathway of the biosynthesis of the branched chain amino acids, valine, leucine and isoleucine. For the first time, the AHAS gene from Bacillus anthracis was cloned into the expression vector pET28a(+), and was expressed in the E. coli strain BL21(DE3). The purified enzyme was checked on 12% SDS-PAGE to be a single band with molecular weight of 65 kDa. The optimum pH and temperature for B. anthracis AHAS was at pH 7.5 and 37 oC, respectively. Kinetic parameters of B. anthracis were as follows: Km for pyruvate, K0.5 for ThDP and Mg2+ was 4.8, 0.28 and 1.16 mM respectively. AHAS from B. anthracis showed strong resistance to three classes of herbicides, Londax (a sulfonylurea), Cadre (an imidazolinone), and TP (a triazolopyrimidine). These results indicated that these herbicides could be used in the search for new anti-bacterial drugs.

Investigation on Bacillus anthracis isolated from Kyong-Ju (경주에서 분리된 탄저균에 대한 연구)

  • 이준규;이은미;차우양;김정화;김영환;이양수;김우현;정종식
    • Korean Journal of Veterinary Service
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    • v.18 no.1
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    • pp.41-56
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    • 1995
  • The present study was conducted to investigate results of B. anthracis isolated from Anthrax in the Kyong-Ju of Feb. 12. 1994. 1. In biochemical feature, B. anthracis was a gram-positive rod, non-motility, sporulation, capsulation. It was positive in gelatinase, starch hydrolysis, glucose. But negative in urease, arabinose, mannitol, xylose. 2. B. anthracis grew well on B4 Br A TSA after incubation for 24 hours. The organisim grew well on BA, Br. A, NA, TSA after incubation for 72 hours. The media grew well on Br A instead of BA. 3. On 5% blood agar by laboratory animal, ${\beta}$ -hemolysis was produced from 36 hours to 48 hours incubation. There was perfect ${\beta}$-hemolysis after incubation for 48 hours. On the other side ${\beta}$-hemolysis was begun on 5% goat blood agar after incubation for 60 hours. 4. In the test of antimicrobial susceptibility, B. anthracis was very sensitive to AM, CF, TE, ENR, GM, AN, DFX, S, P, TYLO, N, KM, C, E, Lins+Sp, NN, CC, CFP, CB were sensitive one by one. B. anthracis was no-sensitive to L, XNL, TIA, CL, SXT 5. B. anthracis had never sensitivity to direct inoculation of rat and chicken, after subcutanous inj. It was very sensitive to mouse and goat, hamster, guinea pig, rabbit had a sensibility one by one. 6. The dead laboratory animal which had been inoculated with B. anthracis preserved at $37^{\circ}C$ incubation, B. anthracis didn't cultivate on non-dissected animal after 80 hours but cultivate on dissected animal after 360 hours. 7. The rapidly death could cause high concentration, died from 420 after S. C. 8. The blood smeared samples of hamster from inoculation with B. anthracis, spore germinated In 37$^{\circ}C$ after 5 hours, in $32^{\circ}C$ after 6 hours, in room temperature after 9 hours, in $-4^{\circ}C$ to $-20^{\circ}C$ after 10 hours. 9. B, anthracis inoculated to laboratory animal after SC or PO. Mice and rats feces didn't cultivated with B. anthracis after SC, but did cultivated with B. anthracis after PO. 10. In the test of disinfectant, B. anthracis was high effective to $HgC1_2$, formalin, effect phenol, cresol, but non-effect NaOH, ethanol.

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Antimicrobial Activities of Hydroxybiphenyl Derivatives (I)

  • Bae, Ki-Hwan;Yoo, Beong-Tae;Lee, Myung-Koo;Seo, Won-Jun
    • Archives of Pharmacal Research
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    • v.8 no.2
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    • pp.85-89
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    • 1985
  • It was revealed that magnolol and honokiol isolated from the stem bark of Magnolia obovata, had potent antibacterial activity against Bacillus anthracis. A quantitative analytical method of magnolol and honokiol by HPLC has been established, and the amounts of the two components in the dried stem bark of M. obovata were 1, 94% and 0.44%, respectively.

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Production of nitric oxide, interleukin-6 and tumor necrosis factor α from mouse peritoneal macrophages in response to Bacillus anthracis antigens

  • Yoo, Han-sang;Kim, Jae-wook;Cho, Yun-sang
    • Korean Journal of Veterinary Research
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    • v.39 no.2
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    • pp.301-310
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    • 1999
  • Anthrax caused by Bacillus anthracis is one of the most important zoonotic diseases. The bacterium produces several virulence factors. Of the factors, protective antigen (PA) of tripatite toxin has been identified as a central component in the pathogenesis of anthrax. However, precise roles of PA and other cellular components in the reaction with the target cells remain to be elucidated, especially in the initial stage of the disease. Three B anthracis antigens were prepared for investigation; PA, sonicated cellular antigens (S-Ag) and formalin-inactivaed whole cell antigens (W-Ag). PA was purified from culture supernatant of the bacterium using FPLC system with MonoQ. S-Ag and W-Ag were prepared by sonication and formalin inactivation of the cultured cells, respectively. Purity of the antigens was confirmed by SDS-PAGE and Western blot analysis. The roles of these antigens in the production of inflammatory mediators such as NO, IL-6 and $TNF{\alpha}$ from mouse peritoneal macrophages were investigated. PA alone did not induce the production of the inflammatory mediators while the other antigens, S-Ag and W-Ag, did in a dose and time dependent manner. These results suggested that in addition to major virulence factors, other cellular antigens are also involved in the initial stage of the disease by the induction of inflammatory mediators.

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Simultaneous Detection and Identification of Bacillus cereus Group Bacteria Using Multiplex PCR

  • Park, Si-Hong;Kim, Hyun-Joong;Kim, Jae-Hwan;Kim, Tae-Woon;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1177-1182
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    • 2007
  • Bacillus cereus group bacteria share a significant degree of genetic similarity. Thus, to differentiate and identify the Bacillus cereus group efficiently, a multiplex PCR method using the gyrB and groEL genes as diagnostic markers is suggested for simultaneous detection. The assay yielded a 400 bp amplicon for the groEL gene from all the B. cereus group bacteria, and a 253 bp amplicon from B. anthracis, 475 bp amplicon from B. cereus, 299 bp amplicon from B. thuringiensis, and 604 bp amplicon from B. mycoides for the gyrB gene. No nonspecific amplicons were observed with the DNA from 29 other pathogenic bacteria. The specificity and sensitivity of the B. cereus group identification using this multiplex PCR assay were evaluated with different kinds of food samples. In conclusion, the proposed multiplex PCR is a reliable, simple, rapid, and efficient method for the simultaneous identification of B. cereus group bacteria from food samples in a single tube.

Protein Expression Profiling of Infected Murine Macrophage Cells (RAW 264.7) by Bacillus anthracis Spores

  • Seo Gwi-Moon;Nam Jeong-Ah;Oh Kwang-Gun;Chai Young-Gyu
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2003.05a
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    • pp.77-79
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    • 2003
  • Current therapeutic strategies far anthrax have had no significant impact on anthrax mortality over the last several decades. This study used a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) discovery platform to generate protein expression profiles in search of overexpressed proteins in murine macrophage cells (RAW264.7) which infected with Bacillus anthracis spores as potentially novel molecular targets. Two differentially expressed proteins were identified in infected murine macrophage cells as Syndapin and CDC46, respectively. Syndapins are potential links between the cortical actin cytoskeleton and endocytosis. Other two proteins were identified from murine macrophage cells infected with avirulent spores as ITBG-2 (CD18) and HSPA5, respectively. These data demonstrate the feasibility of using a MALDI-TOF platform to generate protein expression profiles and identify potential molecular targets for anthrax therapeutics.

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Antimicrobial Activities of Hydroxybiphenyl Derivatives

  • Bae, Ki-Hwan;Koo, Sung-Hyen;Seo, Won-Jun
    • Korean Journal of Pharmacognosy
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    • v.17 no.1
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    • pp.85-90
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    • 1986
  • It has been elucidated that magnolol and honokiol, isolated from the stem bark of Magnolia obovata, had potent antibacterial activity against a cariogenic bacterium Streptococcus mutans. They also show a significant antibacterial activity against Bacillus anthracis, which causes malignant pustule and woolsorter disease in human. Some hydroxybiphenyl derivatives are synthesized from starting materials, phenylphenols and biphenols by means of Claisen's rearrangement and Elb's method to develop more potent antibacterial chemicals and to investigate the structure-activity relationships. The introduction of allyl groups to the aromatic rings of starting materials shows increase of antibacterial activities, but the number and positions of them do not effect their activities. Furthermore, the introduction of hydroxy group to aromatic rings also increases the activity.

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