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Role of the Nfa1 Protein in Pathogenic Naegleria fowleri Co-cultured with Target Cells
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | Shin, Ho-Joon | - |
| dc.contributor.author | 강수연 | - |
| dc.date.accessioned | 2019-10-21T06:45:46Z | - |
| dc.date.available | 2019-10-21T06:45:46Z | - |
| dc.date.issued | 2005 | - |
| dc.identifier.other | 83 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/16305 | - |
| dc.description | 학위논문(석사)--아주대학교 대학원 :의학과,2005 | - |
| dc.description.tableofcontents | TABLE OF CONTENTS ABSTRACT = ⅰ TABLE OF CONTENTS = ⅲ LIST OF FIGURES = ⅴ ABBREVIATIONS = ⅶ Ⅰ. INTRODUCTION = 1 Ⅱ. MATERIALS AND METHODS = 5 A. Cultivation of N. fowleri and CHO cells= 5 B. Expression of the nfa1 gene and production of a rNfa1 protein = 5 C. Production of an anti-Nfa1 polyclonal antibody = 6 D. Immunofluorescent assay and confocal microscopic practice = 7 E. Northern blot analysis = 10 Ⅲ. RESULTS = 11 A. Production of a recombinant Nfa1 fusion protein = 11 B. Characterization of an anti-Nfa1 polyclonal antibody = 12 C. Immunostaining of N. fowleri trophozoites and CHO cells with an anti-Nfa1 polyclonal antibody = 14 D. Counter-staining of N. fowleri trophozoites and CHO cells to observe the localization of the Nfa1 protein = 16 E. Localization of the Nfa1 protein of N. fowleri trophozoites co-cultured with CHO cells = 19 F. Profile of the nfa1 mRNA of N. fowleri in co-culture system = 23 Ⅳ. DISCUSSION = 24 Ⅴ. CONCLUSION = 27 BIBLIOGRAPHY = 28 국문요약 = 32 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | Role of the Nfa1 Protein in Pathogenic Naegleria fowleri Co-cultured with Target Cells | - |
| dc.title.alternative | 표적세포와 혼합배양시 병원성 파울러자유아메바에서 Nfa1 단백질의 역할 | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | 강수연 | - |
| dc.contributor.department | 일반대학원 의학과 | - |
| dc.date.awarded | 2005. 2 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 564450 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000000083 | - |
| dc.description.alternativeAbstract | Naegleria fowleri, a free-living amoeba, exists as a virulent pathogen which causes fatal primary amoebic meningoencephalitis (PAME) in experimental animal and humans. Using infected and immune mouse sera, we previously cloned an antigenic gene (called nfa1) from a cDNA library of N. fowleri by immunoscreening. The nfa1 gene had the coding sequence of 360 bp, producing a 13.1kDa recombinant protein (rNfa1). Anti-Nfa1 polyclonal antibody, revealed pseudopodia-specific immunolocalization of Nfa1 protein in a trophozoite of N. fowleri. An anti-Nfa1 antibody showed a neutralizing effect on the cytotoxicity of N. fowleri trophozoites against CHO cells, much like treating an anti-Nfa1 antibody on a cocultivating system. In spite of the wide use of N. fowleri in free-living amoebic pathogenicity, no informations what proteins are involved in the functions of these organisms are yet available. In this study, we observed the role of Nfa1 protein in a cell-contact mechanism of pathogenic N. fowleri cocultured with target cells (CHO cell) by the immunofluorescence assay. Using confocal microscopic findings, the Nfa1 protein located on pseudopodia of N. fowleri trophozoites. The Nfa1 protein in N. fowleri trophozoites co-cultured with CHO cells was located on pseudopodia and in a food-cup formed as a phagocytic structure close contact with target cells. The amount of nfa1 mRNA of N. fowleri was strongly increased at 6 hr post co-culture. Finally, it was elucidated that the Nfa1 protein played an important role in phagocytic activity, a cell-contact mechanism of pathogenic N. fowleri. | - |
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- Graduate School of Ajou University > Department of Medicine > 3. Theses(Master)
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