2006-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656190摘要：蝦白點病毒(WSSV)已有三株獨立分離之植株且其基因體亦被完全定序，其中台灣分離之WSSV基因體含307 kb及超過500個開放解讀框(open reading frame)。WSSV基因體包含有結構及非結構性蛋白區，其中包括39個以上之結構蛋白及一些非結構性蛋白已被找出並定性，但其功能卻未知。然而過去的努力僅專注於WSSV之診斷及預防，而在蝦白點病毒所引發之宿主反應，我們卻所知不多！因此本計畫之主要目標將專注於研究蝦白點病毒所引發之宿主反應及其調節機制。為完成此一目標，茲簡述逐年計畫如下：在第一年我將建立蝦類細胞初級培養及細胞內鈣離子偵測系統，同時將選殖氧化緊迫及內質網緊迫相關基因。. 在第二年我將(1)純化蛋白質轉譯因子磷酸脢並研究其在WSSV感染期間之活性變化(2)建置WSSV感染期間蝦磷酸化蛋白變化模式圖(3)闡明雙股RNA引發之氧化及內質網緊迫反應(4)利用專一序列之雙股RNA選出可引發之氧化及內質網緊迫反應之病毒蛋白。本計畫之完成將可增進我們對WSSV及宿主之感應機制之瞭解，更重要的是氧化及內質網緊迫反應測試平台之建立可加速我們篩選重要的病毒蛋白以為設計預防及治療WSSV<br> Abstract: The WSSV-genome has been sequenced in three independent isolates. The Taiwanese WSSV isolate contains 307 kb with over 500 open reading frames. It contains both structural and nonstructural proteins. While 39 structural proteins and other viral proteins have been identified and characterized, their functions in eliciting antiviral responses remain mostly unknown. However, despite the effort on the development of diagnosis and prevention of WSSV, our understanding in WSSV-induced responses is still very limited. Therefore, the main objective of this proposal is to investigate the WSSV-induced host responses and their regulatory mechanisms. To achieve the goal, a yearly plan is presented as following: In the first year, I plan to establish the shrimp culture system and have the intracellular calcium measurement platform ready. In addition, I will have the oxidative and ER stress response genes cloned. In the second year I will expect to (1) isolate the shrimp eIF2 kinases and determine its changes in activity upon WSSV infection. (2) collect the change in phosphorylation protein profiles in response toWSSV infection. (3) elucidate the dsRNA-induced oxidative and ER stress responses (4) Pull out WSSV structural proteins capable of triggering intracellular calcium release and antiviral activity. The completion of this project will further our understanding in WSSV and host interaction. More importantly, the establishment of the host response testing platform will speed up our screening of critical viral proteins to be used to devise the preventive and therapeutic strategies for WSSV infection.蝦白點病毒氧化緊迫反應內質網緊迫反應WSSVoxidative stressendoplasmic reticulum stress