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Research on the Entry Mechanism of White Spot Syndrome Virus (Wssv)
|Contributors: ||NTOU:Institute of Marine Biology|
WSSV;PmCBP or CBP;Gluct1;in vitro and in vivo neutralization experiment;WSSV entry model
|Issue Date: ||2013-10-07T02:26:54Z
|Abstract: ||摘要:白點症病毒是造成台灣以及世界其他地區養殖蝦類嚴重死亡的病原體，然而目前 為止我們對於病毒入侵寄主細胞的機制卻仍不清楚。由於蝦類為無脊椎動物，在病毒 性疾病的防治上，最有效的抗病毒策略應為阻斷病毒入侵寄主細胞之機制。在過去的 研究中，我們發現草蝦幾丁質結合蛋白能與白點症病毒封套蛋白VP53A 結合，利用重 組VP53A 和幾丁質結合蛋白對白蝦進行活體感染阻斷試驗，以及對幾丁質結合蛋白基 因進行基因減弱，發現白蝦感染WSSV 後的死亡率可抑制20～50％，顯示幾丁質結合 蛋白與病毒入侵寄主細胞有關。為明白是否有其他蛋白質參與病毒入侵機制，我們利 用酵母菌雙雜合法進行篩選，發現至少有11 個病毒封套蛋白能與幾丁質結合蛋白結 合，也同時發現一新穎蛋白質—葡萄醣運輸蛋白1—可與VP53A 結合，因此我們推測 病毒封套蛋白、寄主細胞膜上幾丁質結合蛋白以及葡萄醣運輸蛋白1 等蛋白質可能形 成一感染複合體，參與了病毒入侵機制。因此在本期的計畫中，我們將由解開病毒封 套蛋白複合體的結構與作用模式著手，冀望能解開白點症病毒入侵寄主細胞機轉，並 選出最好的標的物，再運用口服運送系統以及其他有效方式，開發出最有效的抗病毒 策略，以應用於產業界。|
Abstract:White spot syndrome virus (WSSV) can cause the most serious viral disease of shrimp and has a wide host range among crustaceans. Although researches showed a lot about its genome and structure, information concerning the mechanism of how WSSV infects cells was lacking. Because the shrimp is the invertebrate, the most effective anti-viral strategy is to block the virus entry mechanism. In the previous study, WSSV envelope protein, VP53A, was identified to interact with Penaeus monodon chitin-binding protein (PmCBP, or CBP). In the in vivo infection blocking assay, both rVP53A and rCBP that were produced by Esherichia coli can promote resp. a 40% and 20% survival rate of the shrimp which were challenged by WSSV. Using specific dsRNA to knock-down CBP gene indicated that cumulative mortality of CBP-dsRNA treated group was reduced to 50%. Those findings suggested that CBP was involved in WSSV infection. Furthermore, to reveal whether other proteins participated in the virus entry mechanism, a yeast-two-hybrid experiment was applied and the result indicated that CBP could interact with at least 11 WSSV envelope proteins. Moreover, a new gene, glucose transporter 1 (Gluct1), was also found to interact with VP53A. Thus, we speculate that these 11 WSSV envelope proteins and 2 host proteins may form a complex, infectome, to mediate virus entry. Therefore, a project based on revealing infectome was proposed. A new research working on developing the oral delivery system will also be applied in this project. We hope the study can help to understand the mechanism of virus entry, and the result can ultimately help to select the best candidate to develop the most efficient anti-viral strategy against WSSV.
|Appears in Collections:||[海洋生物研究所] 研究計畫|
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