http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
Multiple-locus Variable-number Tandem Repeat 분석을 사용한 Bacillus Anthracis 균주간 특이성 규명
정경화,김상훈,김성주,김지천,채영규,Jung, Kyoung-Hwa,Kim, Sang-Hoon,Kim, Seong-Joo,Kim, Ji-Cheon,Chai, Young-Gyu 한국군사과학기술학회 2011 한국군사과학기술학회지 Vol.14 No.2
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.
탄저 치사독소 처리에 의한 생쥐 대식세포의 단백질체 발현 양상 분석
정경화,서귀문,김성주,김지천,오선미,오광근,채영규,Jung Kyoung-Hwa,Seo Giw-Moon,Kim Sung-Joo,Kim Ji-Chon,Oh Seon-Mi,Oh Kwang-Geun,Chai Young-Gyu 한국미생물학회 2005 미생물학회지 Vol.41 No.4
탄저 치사독소는 생쥐 대식세포 (RAW 264.7)의 유전자 발현에 많은 변화를 초래한다. 이들 변화를 초래하는 치사독소의 역할은 아직 확실하게 밝혀지지 ???았다. 본 연구에서는, 치사독소가 처리된 생쥐 대식세포의 단백질 프로파일을 이차원 전기영동으로 분석하였고, MALDI-TOF 질량분석기를 사용하여 해당 단백질의 질량을 측정하였다. 펩타이드 질량 분석 데이터는 ProFound 데이터베이스를 이용하여 동정하였다. 차별화되어 발현된 단백질 중에서 절단된 mitogen-activated protein kinase kinase (Mek1)와 glucose-6-phosphate dehydrogenase (G6PD)가 치사독소 처리된 대식세포에서 각각 증가하였다. 치사독소를 처리하였을 경우, Mek1의 절단은 신호전달과정을 방해하고, 증가된 G6PD는 생성된 활성산소로부터 세포를 보호하는 역할을 하는 것으로 보인다. 단백질체 분석기술은 치사독소처리에 의한 생쥐 대식세포의 세포사멸 관련 단백질을 동정하는데 도움을 주어, 치사독소의 잠정적인 기질을 찾는데 유용할 것이다. Intoxication of murine macrophages (RAW 264.7) with the anthrax lethal toxin (LeTx 100 ng/ml) results in profound alterations in the host cell gene expression. The role of LeTx in mediating these effects is unknown, largely due to the difficulty in identifying and assigning function to individual proteins. In this study, we have used two-dimensional polyacrylamide gel electrophoresis to analyze the protein profile of murine macrophages treated with the LeTx, and have coupled this to protein identification using MALDI-TOF mass spectrometry. Interpretation of the peptide mass fingerprint data has relied primarily on the ProFound database. Among the differentially expressed spots, cleaved mitogen-activated protein kinase kinase (Mek1) and glucose-6-phosphate dehydrogenase were increased in the LeTx treated macrophages. Mek1 acts as a negative element in the signal transduction pathway, and G6PD plays the role for the protection of the cells from the hyper-production of active oxygen. Our results suggest that this proteomic approach is a useful tool to study protein expression in intoxicated macrophages and will contribute to the identification of a putative substrate for LeTx.
면역체 분석을 위한 탄저균 유전자 발현 라이브러리의 구축
박문규,정경화,김연희,이기은,채영규,윤장원,Park, Moon-Kyoo,Jung, Kyoung-Hwa,Kim, Yeon-Hee,Rhie, Gi-Eun,Chai, Young-Gyu,Yoon, Jang-W. 한국미생물학회 2010 미생물학회지 Vol.46 No.1
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. As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.
과산화수소 증기를 이용한 다양한 쿠폰 표면의 Geobacillus Stearothermophilus 아포 제독조건
김상훈,정경화,김세계,채영규,김윤기,황현철,김민철,박명규,류삼곤,Kim, Sang Hoon,Jung, Kyoung Hwa,Kim, Se Kye,Chai, Young Gyu,Kim, Yun Ki,Hwang, Hyun Chul,Kim, Min Cheol,Park, Myung Kyu,Ryu, Sam Gon 한국군사과학기술학회 2013 한국군사과학기술학회지 Vol.16 No.4
Biological decontamination means the removal of microorganisms from the inanimate object such as building or equipment. In this study, hydrogen peroxide vapor efficacy test using VHP 1000ED system(Steris LifeSciences) were conducted for G. stearothermophilus spore with agent materials(aluminum, stainless steel, poly-carbonate, viton, silicone, kapton and glass). Total recovered spores exposed to hydrogen peroxide vapor(1.0 g/min) during 7, 15, 30, 60 min were calculated. As a result, all agent materials were totally decontaminated within 60 min at 1.0 g/min concentration with 35% hydrogen peroxide vapor. Finally, we could confirmed that hydrogen peroxide vapor possess sporicidal capacity of G. stearothermophilus and found the optimum decontamination conditions with VHP1000ED system.
장기간 플루세틴 처리에 의한 흰쥐 해마에서의 NCAM140 유전자 발현의 증가
최미란,채영규,정경화,백승연,김석현,노성원,최준호,이준석,최인근,양병환,Choi, Mi Ran,Chai, Young Gyu,Jung, Kyoung Hwa,Baik, Seung Youn,Kim, Seok Hyeon,Roh, Sungwon,Choi, Joonho,Lee, Jun-Seok,Choi, Ihn Geun,Yang, Byung-Hwan 대한생물정신의학회 2009 생물정신의학 Vol.16 No.1
Objectives : Most of the mechanisms reported for antidepressant drugs are the enhancement of neurite outgrowth and neuronal survival in the rat hippocampus. Neural cell adhesion molecule 140(NCAM140) has been implicated as having a role in cell-cell adhesion, neurite outgrowth, and synaptic plasticity. In this report, we have performed to elucidate a correlation among chronic antidepressant treatments, NCAM140 expression, and activation of phosphorylated cyclicAMP responsive element binding protein(pCREB) which is a downstream molecule of NCAM140-mediated intracellular signaling pathway in the rat hippocampus. Methods : Fluoxetine(10mg/kg) was injected acutely(daily injection for 5days) or chronically(daily injection for 14days) in adult rats. RNA and protein were extracted from the rat hippocampus, respectively. Real-time RT-PCR was performed to analyze the expression pattern of NCAM140 gene and western blot analyses for the activation of the phosphorylation ratio of CREB. Results : Chronic fluoxetine treatments increased NCAM140 expression 1.3 times higher than control in rat hippocampus. pCREB immunoreactivity in the rat hippocampus with chronic fluoxetine treatment was increased 4.0 times higher than that of control. Conclusion : Chronic fluoxetine treatment increased NCAM140 expression and pCREB activity in the rat hippocampus. Our data suggest that NCAM140 and pCREB may play a role in the clinical efficacy of antidepressants promoting the neurite outgrowth and neuronal survival.
김상훈,김세계,정경화,윤성녀,김윤기,김민철,류삼곤,이해완,채영규,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 한국군사과학기술학회 2015 한국군사과학기술학회지 Vol.18 No.4
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.