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標題: | 神經壞死病毒外套蛋白與組蛋白H3.3在石斑魚腦細胞株中之交互作用 Interaction of nervous necrosis virus coat protein with histone H3.3 in grouper brain cells |
作者: | Bo-Ting Hsu 許博婷 |
指導教授: | 張繼堯(Chi-Yao Chang) |
關鍵字: | 組蛋白H3.3,神經壞死病毒, Histone H3.3,nervous necrosis virus, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 神經壞死病毒為一種魚類乙型野田病毒,可廣泛感染海水硬骨魚類,造成魚苗和未成熟魚隻嚴重的病毒性神經壞死症和病毒性腦及視網膜病變。本實驗利用病毒覆蓋蛋白結合分析法來探討神經壞死病毒的感染機制,結果鑑定出點帶石斑魚腦細胞株的細胞核蛋白,組蛋白H3.3,會與神經壞死病毒顆粒結合。組蛋白H3.3為組蛋白H3的變異體之ㄧ,此蛋白與組蛋白H2A、H2B以及H4組成一個八聚體的結構提供DNA纏繞形成核小體。點帶石斑魚的組蛋白H3.3含有136個胺基酸,其分子量為15,327 Da,與其他物種的組蛋白H3.3具有極高序列相似性。本實驗利用遠西墨點法確認點帶石斑魚組蛋白H3.3和神經壞死病毒外套蛋白會結合。此結合區域位於點帶石斑魚組蛋白H3.3的35-97胺基酸片段與神經壞死病毒外套蛋白的195-338 胺基酸片段。利用免疫細胞染色法在病毒感染或神經壞死病毒外套蛋白基因轉染的點帶石斑魚腦細胞中,可以觀察到組蛋白H3.3和神經壞死病毒外套蛋白共同座落在細胞中。神經壞死病毒外套蛋白的進入細胞核以及會和組蛋白H3.3結合的特性,暗示神經壞死病毒外套蛋白在病毒感染石斑魚腦細胞後可能扮演一個調控宿主基因表現的角色。 Nervous necrosis virus (NNV), a piscine betanodavirus, is the causing agent of viral nervous necrosis and viral encephalopathy and retinopathy in larvae and juveniles of a wide range of marine teleost species. The mechanism of NNV infection was investigated using a virus overlay protein binding assay (VOPBA), which resulted in the identification of a nuclear protein, histone H3.3, from grouper brain cells. The histone H3.3 is the variant of histone H3 which gets together with H2A, H2B and H4 to form a histone octamer for DNA wrapped binding in nucleosome. The orange-spotted grouper histone H3.3 gene encodes a protein of 136 amino acids with a molecular mass of 15,327 Daltons, and shows higher similarity to other species histone H3.3. The binding of orange-spotted grouper histone H3.3 and NNV coat protein was confirmed by Far-western blotting. The binding regions of orange-spotted grouper histone H3.3 and NNV coat protein were identified to 35 to 97 and 195 to 338 amino acids, respectively. The co-localization of histone H3.3 and NNV coat protein was observed on grouper brain cells through NNV infection and NNV coat protein gene transfection by immunocytostaining. The nuclear entry of NNV coat protein and binding between histone H3.3 and NNV coat protein suggest that NNV coat protein may serves as a modulator to control the expression of host genes post NNV infection in grouper brain cells. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/67182 |
DOI: | 10.6342/NTU201702786 |
全文授權: | 有償授權 |
顯示於系所單位: | 漁業科學研究所 |
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