白點症病毒非結構性蛋白質WSSV071與蝦轉麩醯胺酸酶交互作用關係之探討

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Title:	白點症病毒非結構性蛋白質WSSV071與蝦轉麩醯胺酸酶交互作用關係之探討 Interaction between White Spot Syndrome Virus Nonstructural Protein WSSV071 and Shrimp Transglutaminase
Authors:	張博淵
Contributors:	生物科技系田乃月
Keywords:	酵母菌雙雜交法白點症病毒 WSSV072 蛋白質轉麩醯胺酸酶 Yeast-two hybrid system WSSV072 protein Transglutaminase
Date:	2013
Issue Date:	2014-03-10 20:40:54 (UTC+8)
Abstract:	白點症病毒 ( white pot syndrome virus, WSSV ) 為一種新穎大型的雙股 DNA 病毒，感染水產甲殼類動物，造成養蝦業者嚴重災情與損失。已知 WSSV台灣分離株基因體長有 307,287 bp，雙股 DNA 經雙向核苷酸分析約有 532個開放譯讀區 ( open reading frame, ORFs ) ，目前研究已知部分 ORFs 可譯讀出 WSSV 的套膜、殼膜區間或核殼體等結構性蛋白及部分非結構性蛋白質，其餘則尚有許多未知表現性與功能性之 ORFs，仍待研究分析。在感染繁殖過程中病毒的非結構性蛋白質會短暫於宿主細胞中啟動表現，作用完成後即不存在，其表現的時間性及產量狀況較難輕易掌握。本研究乃探討WSSV 台灣分離株 ( Taiwan isolate, WSSV-TW ) 其中編號為“wssv071”的 ORF 片段；表現性主要利用 real-time PCR 檢測 wssv071 在病毒感染週期中表現時機，初步推測 wssv071 是屬於一個晚期表現的基因，WSSV071 蛋白質功能性質以酵母菌雙雜交法 ( yeast-two hybrid system, Y2H )分析是否與蝦體或病毒本身蛋白質有產生交互作用關係。目前發現WSSV071 蛋白質與完整的轉麩醯胺酸酶 ( transglutaminase, TGase )蛋白質無明顯的交互作用，但其與 TGase 蛋白質的 N 端與 C 端多胜肽片段則似有交互作用關係。 WSSV is a new large double-stranded DNA virus that infects aquatic crustaceans, and causes damages and losses to cultured shrimp. The around 300-kb genome of WSSV Taiwan isolate ( WSSV-TW ) encodes 532 putative open reading frames ( ORFs ). Some of them have been characterized encoding structural proteins of WSSV virions and many nonstructural proteins transiently expression during virus infection. It is study, time-course study using by real-time PCR showed that the wssv071 ORF of WSSV-TW was expressed after 24 hours of infection. The result suggested that wssv071 might be a late gene. Furthermore, Yeast-two hybrid analysis showed that WSSV071 protein might interact with the N-terminal and C-terminal peptide fragments of shrimp transglutaminase ( TGase ).
Relation:	電子全文公開日期：20180901，學年度：101，76頁
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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