The Prevention and Control of Wssv: Development of Oral Delivery System for Virus Entry Inhibitor

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Project title

Code/計畫編號

NSC100-2324-B019-002-CC2

Translated Name/計畫中文名

蝦白點症預防及控制 - 病毒入侵抑制物口服運送系統之開發計畫( I )

Project Coordinator/計畫主持人

Li-Li Chen

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Institute of Marine Biology

Website

https://www.grb.gov.tw/search/planDetail?id=2220870

Year

2011

Start date/計畫起

01-01-2011

Expected Completion/計畫迄

31-12-2011

Bugetid/研究經費

1600千元

ResearchField/研究領域

生物技術(農)
漁業

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，儘管目前在白點症病毒的基因體和蛋白質體學方面的研究相當多，然而對於如何預防及控制此病毒仍無具體策略。在過去的研究中，我們利用酵母菌雙雜合法篩選能與白點症病毒封套蛋白VP53A 結合的蛋白質，發現草蝦幾丁質結合蛋白能與之結合，並驗證確定草蝦幾丁質結合蛋白位在細胞膜上，分子量約為34 kDa。我們利用重組VP53A 和幾丁質結合蛋白對白蝦進行活體感染阻斷試驗，發現白蝦的死亡率可有效抑制，但卻無法百分之百預防，可能是運送至蝦體內的重組蛋白遭酵素破壞，顯示穩定的運送系統有其重要性。最新的研究中中國研究者試圖以枯草桿菌為蛋白質表現載體系統，並直接以表現重組蛋白後的死菌餵食實驗蝦，發現效果較以大腸桿菌為蛋白質表現載體系統為佳，顯示利用革蘭氏陽性菌可能較陰性菌具效果。近來人類癌症治療的研究中使用單核球增多性李斯特菌，利用基因工程技術使此李斯特菌失去致病力卻仍保有其優點，顯示此系統具水產養殖應用之潛力。本計畫將利用已知的VP53A 和幾丁質結合蛋白結合模式為基礎，利用李斯特菌系統，以建立一有效的口服運送系統。"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, the strategies about how to control and prevent this virus were lacking. In the previous study, WSSV envelope protein, VP53A, was identified to interact with Penaeus monodon chitin-binding protein (PmCBP). PmCBP was confirmed to locate on the cell surface of the host cell, and its molecular weight was about 34 kDa. In the in vivo infection blocking assay, both rVP53A and rPmCBP that were produced by Esherichia coli can promote the survival rate of the shrimp which were challenged by WSSV. The neutralization of both rVP53A and rPmCBP significantly, but incompletely, blocked the WSSV infection. This may due to protein degradation without any protection system. In the recent study using Bacillus subtilis as protein production and delivery material showed the better protection activity then using E. coli. It indicated that Gram+ bacteria may be a better protein production and delivery material then Gram- bacteria. Recently, Listeria monocytogenes was applied to human cancer treatment. The pathogenicity of L. monocytogenes was deleted and this system could be attempted to aqualculture. In this study, we plan to develop an effective oral delivery system based on protein-protein interaction between rVP53A and rPmCBP and the newly application of L. monocytogenes. We hope the study can ultimately help to develop the most efficient anti-viral strategy against WSSV."

Keyword(s)

白點症病毒
封套蛋白
草蝦幾丁質結合蛋白
單核球增多性李斯特菌
口服運送系統
WSSV
envelope protein
PmCBP
Listeria monocytogenes
oral delivery system

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The Prevention and Control of Wssv: Development of Oral Delivery System for Virus Entry Inhibitor

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