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

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簡歷

基本資料

Project title

Code/計畫編號

NSC101-2324-B019-001-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( II )

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=2397370

Year

2012

Start date/計畫起

01-01-2012

Expected Completion/計畫迄

31-12-2012

Bugetid/研究經費

1600千元

ResearchField/研究領域

漁業
生物技術(農)

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，並利用重組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，然而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。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基因運送之優點，推測此系統具水產養殖應用之潛力。在前一年度的研究中，我們引用李斯特菌於人類醫療研究中的概念進行測試，先以綠螢光蛋白為報導基因進行分析，發現無論是利用活菌或死菌皆可感染蝦血球細胞，並且將死菌餵食蝦苗或豐年蝦，亦可在腸道中偵測到螢光訊號，顯示利用李斯特菌作為運送平台在蝦體上是可行的。因此在本年度計畫中我們將利用前一年度所測試的結果為基礎，結合已知的VP53A 和幾丁質結合蛋白結合模式，實際進行病毒感染實驗，以驗證此病毒入侵抑制物口服運送系統之效力。"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). 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. Recently, Listeria monocytogenes was applied to human cancer treatment. The pathogenicity of L. monocytogenes was deleted but the gene delivery ability was kept. It indicated that this mutant system could be attempted to aqualculture. In the recent study last year, no matter fed with dead or live L. monocytogenes, the fluorescent signals can be detected in shrimp hemocytes using green fluorescent gene (GFP) as reporter. The fluorescent signals can also be detected in the midgut tissue of shrimp larva and artemia after fed with dead L. monocytogenes. The results showed the possibility of inducing L. monocytogenes system for gene delivery in shrimp. In the study this year, we plan to combine the results we got last stage and the concept of protein-protein interaction between VP53A and PmCBP. WSSV envelope proteins will be applied to the experiments to identify the efficiency of our oral delivery system against virus infection."

Keyword(s)

白點症病毒
封套蛋白 VP53A
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 VP53A
PmCBP
Listeria monocytogenes
oral delivery system

DSpace CRIS

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

基本資料

Description