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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/48683

Title: 點帶石斑腦型肌酸激酶與神經壞死病毒交互作用之分析
Analysis of interaction between Epinephelus coicoides creatine kinase brain type and nervous necrosis virus
Authors: Wang, Szu-Wen
王思文
Contributors: NTOU:Institute of Marine Biology
國立臺灣海洋大學:海洋生物研究所
Keywords: 神經壞死病毒;酵母菌雙雜合法;腦型肌酸激酶;遠端西方點墨法;免疫螢光染色
Nervous necrosis virus;yeast two hybrid;creatine kinase brain type;far-western blotting;immunofluorescence staining
Date: 2017
Issue Date: 2018-08-22T06:15:01Z
Abstract: 由於石斑魚具有很高的經濟價值,因此石斑魚養殖產業成為台灣的重要產業之一,然而神經壞死病毒卻對石斑魚養殖產生嚴重的致死率,尤其在石斑魚苗的繁殖培育上,致死率近乎百分之百。為了瞭解神經壞死病毒的感染途徑,實驗室之前利用酵母菌雙雜合法,找出神經壞死病毒殼體蛋白在感染過程中蛋白與蛋白間的交互作用,進而研究對抗神經壞死病毒的對策,而肌酸激酶為候選蛋白之一。 肌酸激酶為生物體內調節能量恆定的酵素之一,其中腦型肌酸激酶蛋白為石斑魚腦部中比例最多的異構型,而神經壞死病毒又是主要感染魚的腦部與視網膜。因此在本研究中將探討腦型肌酸激酶與神經壞死病毒的交互作用。在體外實驗中,從遠端西方點墨法的結果看來,大腸桿菌生產的重組型肌酸激酶可能與重組神經壞死病毒的殼體蛋白交互作用,殼體蛋白為神經壞死病毒外殼唯一的結構蛋白,決定著病毒的宿主範圍。利用細胞轉染將帶有腦型肌酸激酶基因的質體送入GF1細胞株使其表現,再感染神經壞死病毒進行免疫螢光染色,發現在腦型肌酸激酶與神經壞死病毒共同存在於細胞質中。依據實驗結果推測腦型肌酸激酶蛋白可與神經壞死病毒的殼體蛋白交互作用,並在神經壞死病毒感染過程中扮演重要角色。
quaculture of grouper is one of important industries in Taiwan due to its high economic value. However, this industry is damanged by nervous necrosis virus (NNV). NNV can cause mass mortality in grouper aquaculture especially grouper hatchery. Mortality is near 100% in grouper fries and larvae stage. To understand infection pathway of nervous necrosis virus, the yeast two hybrid screen was performed to discover protein-protein interaction between NNV coat protein and its host. Creatine kinase (CK) is one of candidate proteins that may interact with nervous necrosis virus.   CK is a kind of enzyme for maintenance of energy homeostasis. Creatine kinase brain type (CKB), one of CK isoforms, mainly exists in brain organ. On the other hand, brain is also main infection location of NNV. Therefore, analysis of interaction between CKB and NNV is key point in this study. In vitro experiment showed that CKB can bind NNV coat protein (NNV CP). Besides, CKB showed colocalize with NNV CP in GF1 cell. Thus, CKB is speculated that it can interact with NNV CP and plays important in NNV infection.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010434006.id
http://ntour.ntou.edu.tw:8080/ir/handle/987654321/48683
Appears in Collections:[海洋生物研究所] 博碩士論文

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