• 제목/요약/키워드: AMOEBA

검색결과 104건 처리시간 0.039초

Development of a Plasmid Vector for Overproduction of $\beta$-Galactosidase in Escherichia coli by Using Genetic Components of groEx from Symbiotic Bacteria in Amoeba proteus

  • Lee, Jung-Eun;Ahn, Eun-Young;Ahn, Tae-In
    • Journal of Microbiology and Biotechnology
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    • 제8권5호
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    • pp.509-516
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    • 1998
  • A plasmid vector, pXGPRMATG-lac-Tgx, was developed for overproduction of $\beta$-galactosidase in Escherichia coli using the genetic components of groEx, a heat-shock gene cloned from symbiotic X-bacteria in Amoeba proteus. The vector is composed of intragenic promoters P3 and P4 of groEx, the structural gene of lac operon, transcription tenninator signals of lac and groEx, and ColEl and amp'of pBluescript SKII. The optimized host, E. coli DH5$\alpha$, transfonned with the vector constitutively produced 117,310-171,961 Miller units of $\beta$-galactosidase per mg protein in crude extract. The amount of enzyme in crude extract was 53% of total water-soluble proteins. About 43% of the enzyme could be purified to a specific activity of 322,249 Miller units/mg protein after two-fold purification, using two cycles of precipitation with ammonium sulfate and one step of gel filtration. Thus, the expression system developed in this study presents a low-cost and simple method for purifying overproduced $\beta$-galactosidase in E. coli.

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Down-Regulation of Cellulose Synthase Inhibits the Formation of Endocysts in Acanthamoeba

  • Moon, Eun-Kyung;Hong, Yeonchul;Chung, Dong-Il;Goo, Youn-Kyoung;Kong, Hyun-Hee
    • Parasites, Hosts and Diseases
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    • 제52권2호
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    • pp.131-135
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    • 2014
  • Acanthamoeba cysts are resistant to unfavorable physiological conditions and various disinfectants. Acanthamoeba cysts have 2 walls containing various sugar moieties, and in particular, one third of the inner wall is composed of cellulose. In this study, it has been shown that down-regulation of cellulose synthase by small interfering RNA (siRNA) significantly inhibits the formation of mature Acanthamoeba castellanii cysts. Calcofluor white staining and transmission electron microscopy revealed that siRNA transfected amoeba failed to form an inner wall during encystation and thus are likely to be more vulnerable. In addition, the expression of xylose isomerase, which is involved in cyst wall formation, was not altered in cellulose synthase down-regulated amoeba, indicating that cellulose synthase is a crucial factor for inner wall formation by Acanthamoeba during encystation.

22.9[kV]급 애자의 표면오염에 의한 자외선 영상과 열화판정 (The UV Images and Aging Judgement with Behaviors Caused by Surface Contamination on 22.9[kV] Class Insulators)

  • 송길목;김영석
    • 조명전기설비학회논문지
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    • 제24권1호
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    • pp.167-174
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    • 2010
  • 본 연구에서는 22.9[kV] 애자의 표면오염에 의한 자외선 영상과 거동을 통해 자외선 이미지 형태 판단법을 정의하였다. 코로나 방전에 의해 나타나는 자외선 영상은 아메바, 젤리피쉬, 썬플라워 형태 등 3가지로 나누어지며, 세부적으로는 8개의 진행 메커니즘으로 구분하였다. 전력설비가 설치된 현장에서는 즉각적으로 이 판단 방법으로 전력 설비의 열화된 부분을 찾을 수 있다. 이는 신뢰성 있는 데이터를 통해 전력설비의 열화를 판단하는 기준에 활용이 가능하다. 향후 현장진단에 적합한 기술로서의 연구가 활용될 것이다.

Amoeba proteus xD Strain의 변이주 특이성 단백질의 운영 (The Fate of Strain-Specific Protein in xD Strain of Amoeba proteus)

  • 안태인
    • 한국동물학회지
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    • 제26권3호
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    • pp.181-192
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    • 1983
  • 이차원 전긱영동법에 의하여 A. proteus의 tD strain과 tD strain이 박테리아와 공생에 의해 유도된 xD strain의 단백질 양성을 비교하였다. Silver stain에 의해 비교 가능한 200여개의 주요 단백질 가운데 tD strain에서 분자량 45,000, 동전점 5.9의 특이성 단백질이, xD strain의 세포액과 공생낭에서는 분자량 29,000, 동전점 5.5의 공생 특이성 단백질이 탐지되었다. 공생 특이성 단백질은 아메바 고은 배양 및 실험 공생 아메바를 이용한 실험 결과 박테리아와 직접 연관 되어 있었다. 탐지된 두 특이성 단백질에 대하여 종전의 세포 핵 이식 및 배양 실험을 통해서 얻어진 결과에 비추어 이들 단백질 상호 연관 및 세포내의 기능에 관하여 논의하였다.

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Effects of Mannose on Pathogenesis of Acanthamoeba castellanii

  • Yoo, Kyung-Tae;Jung, Suk-Yul
    • Parasites, Hosts and Diseases
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    • 제50권4호
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    • pp.365-369
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    • 2012
  • Acanthamoeba spp. are single-celled protozoan organisms that are widely distributed in the environment. In this study, to understand functional roles of a mannose-binding protein (MBP), Acanthamoeba castellanii was treated with methyl-alpha-D-mannopyranoside (mannose), and adhesion and cytotoxicity of the amoeba were analyzed. In addition, to understand the association of MBP for amoeba phagocytosis, phagocytosis assay was analyzed using non-pathogenic bacterium, Escherichia coli K12. Amoebae treated with mannose for 20 cycles exhibited larger vacuoles occupying the most area of the amoebic cytoplasm in comparison with the control group amoebae and glucose-treated amoebae. Mannose-selected amoebae exhibited lower levels of binding to Chinese hamster ovary (CHO) cells. Exogenous mannose inhibited >50% inhibition of amoebae (control group) binding to CHO cells. Moreover, exogenous mannose inhibited amoebae (i.e., man-treated) binding to CHO cells by <15%. Mannose-selected amoebae exhibited significantly decreased cytotoxicity to CHO cells compared with the control group amoebae, 25.1% vs 92.1%. In phagocytic assay, mannose-selected amoebae exhibited significant decreases in bacterial uptake in comparison with the control group, 0.019% vs 0.03% (P<0.05). Taken together, it is suggested that mannose-selected A. castellanii trophozoites should be severely damaged and do not well interact with a target cell via a lectin of MBP.

The role of domestic tap water in Acanthamoeba contamination in contact lens storage cases in Korea

  • JEONG Hae Jin;YU Hak Sun
    • Parasites, Hosts and Diseases
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    • 제43권2호
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    • pp.47-50
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    • 2005
  • A survey was carried out from August to December 2004 in Pusan, Korea to document the presence of free-living amoeba (FLA), including the genus Acanthamoeba, in both contact lens storage cases and domestic tap water. Acanthamoeba was isolated from $5(4.2\%)$ in 120 contact lens storage cases. Four house tap water samples from residents, whose contact lens storage cases had been contaminated by Acanthamoeba, were also found to be contaminated with Acanthamoeba. Therefore, the contamination rate of FLA and Acanthamoeba in domestic tap water was investigated in order to examine the role of domestic tap water in Acanthamoeba contamination of contact lens storage cases. FLA and Acanthamoeba were identified in $97(46.8\%)\;and\;16(7.7\%)$ of the 207 domestic tap water samples, respectively. There were no significant differences between the contamination rates of FLA in tap water according to the filtration plant of origin. No FLA was detected in the tap water directly supplied by the water purification plants. Water storage tanks appear to promote FLA colonization, including Acanthamoeba, in domestic tap water. This increases the risk of Acanthamoeba contamination in contact lens storage cases as well as increasing the risk of Acanthamoeba keratitis.

Characterization of Ubiquitinated Lysosomal Membrane Proteins in Acanthamieba castellanii

  • Oh, Sekyung;Ahn, Tae-In
    • Animal cells and systems
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    • 제4권2호
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    • pp.165-171
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    • 2000
  • Ubiquitinated proteins in lysosomes were characterized by using two monoclonal antibodies (mAbs): LYS8-1, a mAb to lysosomal proteins, and NYA124, a mAb to ubiquitin. LYS8-1 stained lysosome-like vesicles in immunofluorescence microscopy of Amoeba proteus and Acanthamoeba castellanii. In immunoblotting, LYS8-1's antigens (LYS proteins) were detected as 68-kDa and 77-kDa proteins in A. proteus, and as 30-kDa and 39-kDa proteins in A. castellanii. In immunoprecipitation of A. castellanii, at least four distinct LYS proteins, LVS35p, LyS39p, LyS42p, and LYS46p, were detected and accumulated upon inhibition of lysosome functions but not upon that of 26S proteasome functions. They were all found to be ubiquitinated, and were recovered in the lysosome fractions in subcellular fractionation experiments. In chemical fractionation analyses, LYS35p and LYS39p were demonstrated to be peripherally associated with lysosome membrane, while LYS42p and LYS46p tightly bound to the membrane. These results suggest that the LYS proteins become associated to lysosomal membrane upon ubiquitination.

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