• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.806-811
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• 2007

Bacillus anthracis is a soil pathogen capable of causing anthrax that is closely related to several environmental species, including B. cereus, B. mycoides, and B. thuringiensis. DNA homology studies showed that B. anthracis, B. cereus, B. mycoides, and B. thuringiensis are closely related, with a high sequence homology. To establish a method to specifically detect B. anthracis in situations such as environmental contamination, we initially performed RAPD-PCR with a 10-mer random primer and confirmed the presence of specific PCR bands only in B. anthracis species. One region specific for B. anthracis was cloned and sequenced, and an internal primer set was designed to amplify a 241-bp DNA fragment within the sequenced region. The PCR system involving these specific primer sets has practical applications. Using lyses methods to prepare the samples for PCR, it was possible to quickly amplify the 241-bp DNA segment from samples containing only a few bacteria. Thus, the PCR detection method developed in this study is expected to facilitate the monitoring of environmental B. anthracis contamination.

• 미생물학회지
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• 제37권1호
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• pp.56-60
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• 2001

탄저균의 분자적 다양성 분석은 다양한 DNA표지의 부족으로 쉬운 일이 아니어서, 본 연구에서는 random amplified polymorphic DNA (RAPD)-PCR을 이용하여 Bacillus 속으로부터 탄저균을 구별할 수 있는 새로운 DNA 표지를 개발하고자 하였다. RAPD-PCR을 이용한 분석은 다양한 Bacillus 종으로부터 탄저균을 동정할 수 있었으며, 아울러 Bacillus 종 사이에서 확실한 유전적인 변이를 확인할 수 있었다. 이러한 분석은 간단, 신속하고, 그리고 정확하게 탄저균을 진단하는데 활용할 수 있다고 본다.

• 대한수의학회지
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• 제38권3호
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• pp.574-580
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• 1998

Anthrax caused by Bacillus anthracis is one of the most important zoonotic diseases in the worldwide. To control and prevent the disease effectively, several methods such as development of a fast and specific diagnostic method and vaccine, education etc, have been carried out. However, it still has a problem in the control and prevention. To control, the most important method is the prevention of direct or indirect contact of the causative agent with susceptible host. Therefore, we developed a fast and specific detection method, polymerase chain reaction, of B anthracis from soil and infected animals because the organism could survive long time in the environment including soil due to formation of spore. With the method, virulence genes of B anthracis were successfully amplified from experimentally infected soil and mice. Up to $4.2{\times}10$ of the organisms per gram could be detected with the PCR method from experimentally infected soil. These results suggested that this PCR method could be effectively used not only to detect B anthracis in soil and infected animal but also to provide the information to prevent the disease.

• 한국군사과학기술학회지
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• 제18권4호
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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.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.778-783
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• 2008

Anthrax is an infectious disease caused by toxigenic strains of the Gram-positive bacterium Bacillus anthracis. To identify the mitochondrial proteins that are expressed differently in murine macrophages infected with spores of B. anthracis Sterne, proteomic and MALDI-TOF/MS analyses of uninfected and infected macrophages were conducted. As a result, 13 mitochondrial proteins with different expression patterns were discovered in the infected murine macrophages, and some were identified as ATP5b, NIAP-5, ras-related GTP binding protein B isoform CRAa, along with several unnamed proteins. Among these proteins, ATP5b is related to energy production and cytoskeletal rearrangement, whereas NIAP-5 causes apoptosis of host cells due to binding with caspase-9. Therefore, this paper focused on ATP5b, which was found to be down regulated following infection. The downregulated ATP5b also reduced ATP production in the murine macrophages infected with B. anthracis spores. Consequently, this study represents the first mitochondrial proteome analysis of infected macrophages.

• Bulletin of the Korean Chemical Society
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• 제28권7호
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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.

• 미생물학회지
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• 제37권1호
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• pp.21-27
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• 2001

Bacillus anthracis Sterne 34F$_2$균주로부터 PA를 생산하기 위해 RM배지를 변형하였다. NaHCO$_3$를 8 g/l에서 10 g/l로, glucose를 5 g/l로 첨가하여 새로운 배지조성에서 탄저균을 배양한 후, 배양액을 hydroxyapatite를 이용하여 농축하였다. 농축된 조단백질을 hydroxyapatite column chromatography, DEAE-Sepharose CL-4B column chromatography 및 Toyo-pearl gel filtration chromatography를 사용하여 PA를 정제하였다. 변형된 RM 배지를 사용해 얻은 PA 양은 8.6 mg/l 이었다.

• Archives of Pharmacal Research
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• 제8권2호
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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.

• Journal of Photoscience
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• 제9권2호
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• pp.323-325
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• 2002

Bacillus Anthracis is the causative agent of anthrax. The major virulence factors are a poly-D glutamic acid capsule and three-protein component exotoxin, which is collectively known as anthrax toxin, protective antigen (PA, 83 kDa), lethal factor (LF, 90 kDa), and edema factor (EF, 89 kDa). These three proteins individually have no known toxic activities, but in combination with PA form two toxins (lethal toxin and edema toxin), causing different pathogenic responses in animals and cultured cells. However, it remains to be elucidated for pathogenic mechanism of anthrax toxin. In this study, we constructed toxin component in bacterial overexpression system and purified the native toxin from Bacillus anthracis delta sterne F32 using FPLC system. Recombinant toxin showed high homogeneity and rapid purification processes. Also, this recombinant toxin was comparable to B. anthracis native toxin in terms of cytotoxic effects on cultured cell lines.

• 한국군사과학기술학회지
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• 제14권2호
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• pp.305-312
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• 2011

Bacillus anthracis(Ba) is a Gram-positive spore-forming bacterium that causes the disease anthrax. The feature of Ba is the presence of two large virulence plasmids, pXO1 and pXO2. Molecular genotyping of Ba has been difficult to the lack of polymorphic DNA marker. Ba isolated from Korea has been genotyped using various nucleotide analysis methods, such as 16s rDNA sequencing and multiple-locus variable-number tandem repeat (MLVA) analysis. We identified genotypes that represent a genetic lineage in the B1 cluster. This study emphasized the need to perform molecular genotyping when attempting to verify a strain-specific Ba.