• 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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• 제7권1호
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• pp.77-81
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• 2004

The etiological agent is Bacillus anthracis, a gram-positive rod-shaped bacterium able to form spores. In order to elucidate the mechanism of infecttion on human macrophage cells, we performed two-dimensional electrophoresis and MALDI-TOF analysis using the infected human macrophage cells with the spores of B. anthracis Sterne of inactivated B. anthracis Sterne. We identified 9 proteins which related to the infection of Bacillus anthracis spores on human macrophage cells at the early stage events. Maybe nine proteins will be bio-markers and vaccine candidates to the Bacillus anthracis spore infection.

• 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.

• 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.

• 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.

• 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.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.

• Archives of Pharmacal Research
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• 제15권1호
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• pp.58-61
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• 1992

Bacillus anthracis 590 having an inducibla resistance determinant to MLS antibiotics was isolated from a soli sample in Korea. The resistance gene (ernJ) was cloned by Southern blotting of chromosomal DNA fragment digested by various restriction enzymes and coloy hybridization method and the cloned plasmid was named as pBA423. The size of inserted DNA fragment of pBS42 vector was about 2.9 kb and the DNA sequence of the subcloned fragment (Hinc II-Hinc II, 1.4kb) WAS determined. The DNA sequence of ernJ was composed of 357 bp for leader region and 861 bp for the structural gene. Because the leader sequence of ernJ was homologous to that of ermK, the expression of ernJ is also thought to be controlled by a transcriptionl attenuation mechanism.

• 미생물학회지
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• 제46권1호
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• pp.21-26
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• 2010

탄저균(Bacillus anthracis)은 탄저(Antrax)의 원인균으로 사람은 물론, 초식동물인 소, 양, 말 등에서 급성의 폐사성 전염병을 일으킨다. 현재 사용되고 있는 탄저 치료 및 예방법은 항생제 치료와 약독화 백신주를 토대로 하고 있으나, 항생제 내성주의 출현 및 잔류 병원성이 문제시 되고 있는 실정이다. 따라서, 인체에 적용 가능하며 보다 안전한 탄저 치료제 및 백신 개발이 요구되고 있으며, 최근 탄저균 아포 및 영양세포, 그리고 탄저독소(Anthrax toxins)에 대한 동시 면역을 유도하는 다가백신 개발이 보고된 바 있다. 본 연구에서는, 향후 탄저균에 대한 새로운 다가백신 후보물질 발굴을 위하여, 탄저균에 대한 전장 유전자 발현 라이브러리(whole genomic expression library)를 구축하였다. 라이브러리 구축을 위하여, 탄저균(ATCC 14578) 게놈 DNA를 Sau3AI으로 부분 제한효소 처리였고, 유도 발현이 가능한 pET30abc 벡터에 접합시킴으로써, 총 $1{\times}10^5$개에 해당하는 대장균 BL21(DE3) 유래의 전장 유전자 발현 라이브러리를 구축하였다. 염기서열분석을 통한 중복성(redundancy) 확인 결과, 111개의 무작위 클론 중 56개(50.5%)가 탄저균 유전자로 확인되었으며, 17개(15.3%)는 벡터 유전자였고, 38개(34.2%)는 BLAST 탐색에서 일치하는 유전자를 찾지 못하였다. 또한 웨스턴 분석을 통하여 단백질 유도발현을 확인하였으며, 탄저균 항혈청에 대한 colony blot으로부터 양성반응을 보이는 일부 클론들을 확인할 수 있었다. 이러한 결과물들은, 구축된 전장 유전자 발현 라이브러리가 향후 탄저균에 대한 면역체(immunome) 분석을 위해 적용 가능함을 암시한다.

• 셀메드
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• 제4권1호
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• pp.6.1-6.12
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• 2014

Anthrax is the deadly disease for human being caused by Bacillus anthracis. Instantaneous research work on the mode of infection of the organism revealed that different proteases are involved in different steps of pathogenesis. Present study reports the in silico characterization and the detection of pathogenic proteases involved in anthrax infection through protein-protein interaction. A total of 13 acid, 9 neutral, and 1 alkaline protease of Bacillus anthracis were selected for analysing the physicochemical parameter, the protein superfamily and family search, multiple sequence alignment, phylogenetic tree construction, protein-protein interactions and motif finding. Among the 13 acid proteases, 10 were found as extracellular enzymes that interact with immune inhibitor A (InhA) and help the organism to cross the blood brain barrier during the process of infection. Multiple sequence alignment of above acid proteases revealed the position 368, 489, and 498-contained 100% conserved amino acids which could be used to deactivate the protease. Among the groups analyzed, only acid protease were found to interact with InhA, which indicated that metalloproteases of acid protease group have the capability to develop pathogenesis during B. anthracis infection. Deactivation of conserved amino acid position of germination protease can stop the sporulation and germination of B anthracis cell. The detailed interaction study of neutral and alkaline proteases could also be helpful to design the interaction network for the better understanding of anthrax disease.