碩士 / 國立臺灣海洋大學 / 海洋生物研究所 / 100 / White spot syndrome virus (WSSV) has caused serious economic impact on global shrimp aquaculture industry. Although researches showed a lot about its genome and structure, information concerning the mechanism of how WSSV infects cells is lacking. Our previous research indicated that a shrimp surface protein, chitin binding protein (CBP), is a crucial protein in WSSV infection. In this study, observation the interaction between CBP and WSSV on model cells and identification the process of WSSV entry are the main focuses in this study. The Spodoptera frugiperda Sf9 cell line was used as an analysis platform to identify the interaction of CBP and WSSV. The immunofluorescent microscopy observation showed that more WSSV signals were found on the surface of the cell transfected CBP. Furthermore, primary cultured hemocytes were used as an experimental model to observe the process of WSSV entry because the stable shrimp cell lines for WSSV infection are lacking. Our results show that the WSSV colocalized with early endosome. By using endocytic inhibitor methyl-β-cyclodextrin (MβCD) and chlorpromazine (CPZ), WSSV still can be detected in the hemocytes treated with CPZ, but not in the hemocytes treated with MβCD. Thus, we defined that WSSV enter shrimp cell by binding with CBP, and then adopts the endocytosis pathway with cholesterol assistance.
| Identifer | oai:union.ndltd.org:TW/100NTOU5270013 |
| Date | January 2012 |
| Creators | Zih-Jhan Huang, 黃子禎 |
| Contributors | Li-Li Chen, 陳歷歷 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 57 |
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