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研究生:吳雅惠
研究生(外文):Wu Ya-Hui
論文名稱:豬生殖與呼吸綜合症病毒ORF3基因之表現
論文名稱(外文):Expression of ORF3 Gene of Porcine Reproductive and Respiratory Syndrome Virus
指導教授:闕玲玲
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:獸醫學研究所
學門:獸醫學門
學類:獸醫學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:英文
論文頁數:68
中文關鍵詞:豬生殖與呼吸綜合症病毒ORF3基因綠色螢光蛋白表現
外文關鍵詞:porcine reproductive and respiratory syndrome virusORF3green fluorescent proteinexpression
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為了解豬生殖與呼吸綜合症病毒ORF3蛋白之表現及性質,本研究將台灣分離株MD-001之ORF3基因構築於真核綠色螢光蛋白表現載體,以便追蹤ORF3綠色螢光融合蛋白的表現。結果顯示實驗組之ORF3綠色螢光蛋白只出現在細胞質,並有聚集在細胞一端的情況;而控制組之綠色螢光蛋白則均勻分佈於整個細胞。此外,實驗組表現之綠色螢光的細胞數量雖然不多,但其表現細胞卻多呈圓形化。以西方轉漬法可確認綠色螢光蛋白在控制組細胞內的表現,但是不管在細胞內或培養的上清液中皆無法偵測到ORF3綠色螢光蛋白。為瞭解此結果是否與表現量過低有關,進一步分析兩組細胞表現的綠色螢光蛋白mRNA,發現實驗組與控制組的基因表現量無明顯差異。未來需要設計不同實驗釐清上述現象是否為ORF3蛋白可能具有細胞毒性相關特性或是與綠色螢光蛋白融合所致,以期進一步研究ORF3蛋白的功能。

To validate the character of the GP3 of porcine reproductive and respiratory syndrome virus (PRRSV), full-length of open reading frame 3 (ORF3) gene of Taiwanese strain MD-001 was fused with eukaryotic green fluorescent protein (GFP) expression vector. The results showed that GFP/ORF3 fusion protein was localized mainly in the cytoplasm and then accumulated progressively at one-side of cells, whereas the GFP distributed throughout the whole expressing cells. The amount of GFP/ORF3-expressing cells was rare while most of them were rounded-up. Western blot analysis did not allow the identification of the GFP/ORF3 in both fractions of cell pellets and cell culture medium, but RNA analysis indicated the amount of gene expression in two constructions was similar. Rapid degradation of misfolded GFP/ORF3 or cell toxicity of GFP/ORF3 might interfere with the fate of GP3, and further experiments will be needed to clarity these questions.

Abstract Ⅰ
中文摘要 Ⅱ
Table of contents Ⅲ
List of figures Ⅳ
Chapter Ⅰ Introduction 1
Chapter Ⅱ Introduction of the porcine reproductive and respiratory syndrome virus 2
2-1 The history and epidemiology 2
2-2 The taxonomy of PRRSV 4
2-3 The viral structure and characteristics 5
2-4 The cell receptors of PRRSV 7
2-5 The genome properties and organization of PRRSV 10
2-6 The structural proteins of PRRSV 11
2-7 The GP3 protein of PRRSV 14
Chapter Ⅲ Materials and methods……….……………………18
Chapter Ⅳ Results…………………………...…....…………36
Chapter Ⅴ Discussion ………………………………………….55
References …………………………………………………......59

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