English  |  正體中文  |  简体中文  |  Items with full text/Total items : 26988/38789
Visitors : 2341990      Online Users : 42
RC Version 4.0 © Powered By DSPACE, MIT. Enhanced by NTU Library IR team.
Scope Adv. Search
LoginUploadHelpAboutAdminister

Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/16898

Title: 點帶石斑魚抗病毒基因viperin 分子選殖及其特性分析
Molecular cloning and characterization of viperin gene in orange-spotted grouper (Epinephelus coioides)
Authors: Jr-shiang Yang
楊智翔
Contributors: NTOU:Department of Aquaculture
國立臺灣海洋大學:水產養殖學系
Keywords: 點帶石斑;抗病毒基因;先天性免疫;中草藥;神經壞死病毒
viperin;Poly I:C;LPS;peptidoglycan;beta-1,3-glucan
Date: 2008
Issue Date: 2011-06-30T08:44:08Z
Abstract: 石斑魚為一種高經濟養殖魚種,但由於近年來病毒性疾病爆發,造成經濟上大規模損失,石斑魚的疾病防治成了一個重要的議題。Viperin基因為一個先天性免疫因子,可被不同形式干擾素與病毒所誘導表現且被證實具有抗病毒之活性。本研究目的是以分子生物學技術研究點帶石斑魚viperin基因生化特性及其抗病毒活性。利用已發表之其他物種viperin核酸與胺基酸序列,設計退化性引子。以點帶石斑頭腎之mRNA使用RT-PCR及RACE方法選殖出viperin基因序列,其全長為1367個核苷酸,可轉譯出361個胺基酸。經由即時定量聚合酶連鎖反應分析viperin基因各組織表現量,結果顯示點帶石斑viperin基因廣泛地表現於各組織當中,其中又以頭腎與脾臟的表現量為最高。注射Poly I:C後viperin基因在眼睛、腦與脾臟中表現量均有顯著增加,且均隨著時間增加而增加表現量;注射免疫刺激物LPS後,viperin基因表現量在鰓、頭腎與脾臟中均有顯著增加,且均隨著時間增加而增加表現量。而當點帶石斑感染神經壞死病毒 (NNV) 後,血液、腦、鰓、頭腎、脾臟、眼睛、鰭與心臟等組織中viperin基因表現量皆較未感染魚隻的來得高,且以血液與腦組織的表現量為最高。在點帶石斑魚苗發育時期中viperin基因表現量在2.3吋時期的表現量較石斑魚苗2吋的表現量來得高,顯示內生型免疫系統在2吋前的石斑魚苗中不完全發育。以其他免疫刺激物進行注射,發現魚苗時期viperin基因可被peptidoglycan所誘導;而中草藥處理實驗中則發現,甘草、金銀花、魚腥草可誘導viperin基因表現。抗病毒能力實驗中發現,甘草處理組可以增加經神經壞死病毒感染後魚苗之活存率。
Grouper is one of the most important aquacultured fish in Taiwan and disease prevention is a vital issue to grouper culturing as infectious viral diseases had caused great damages in grouper aquaculture. It has been demonstrated that viperin can be induced by type-I/II interferon and viruses, its antiviral activities also be proved. The goal of present study is cloning and characterization of viperin gene in orange-spotted grouper (Epinephelus coioides) and study its antiviral activity. A degenerated primers were designed base on known viperin amino acid and nucleic acids sequences. A full length grouper viperin gene cDNA is isolated from head kidney by using RT-PCR and 5’ and 3’ RACE techniques. Viperin gene cDNA consists of 1367 base pairs and can be deduced into 361 amino acids. Viperin gene mRNA was detected in multiple tissues, and highest mRNA level was detected in head kidney and spleen. Furthermore, dsRNA Poly I:C injections can induce viperin gene expression in eye, brain and spleen tissues of grouper, and with a time effect manner. Microbial LPS injections also can induce viperin gene expression in gill, spleen and head kidney tissues of grouper, and with a time effect manner. After infecting with NNV, higher expression level of viperin gene were detected in gill, head kidney, spleen, eye, fin and heart tissues, the highest level of viperin mRNA was detected in blood and brain. During grouper larval development stages, the viperin mRNA was increased significantly after the stage of 2.3 inch in length. Viperin gene expression arose significantly by injection of peptidoglycan and also by Chinese herbs treatment such as Glycyrrhiza glabra, Lonicera japonica and Houttuynia cordata. The result of NNV challenge test indicated that Glycyrrhiza glabra treatment could improve the survival rate of orange-spotted grouper following NNV infections.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0M95330021
http://ntour.ntou.edu.tw/ir/handle/987654321/16898
Appears in Collections:[水產養殖學系] 博碩士論文

Files in This Item:

File Description SizeFormat
index.html0KbHTML159View/Open


All items in NTOUR are protected by copyright, with all rights reserved.

 


著作權政策宣告: 本網站之內容為國立臺灣海洋大學所收錄之機構典藏,無償提供學術研究與公眾教育等公益性使用,請合理使用本網站之內容,以尊重著作權人之權益。
網站維護: 海大圖資處 圖書系統組
DSpace Software Copyright © 2002-2004  MIT &  Hewlett-Packard  /   Enhanced by   NTU Library IR team Copyright ©   - Feedback