Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/52609
標題: 以螢光標示融合蛋白鑑別診斷豬瘟抗體之研究
Study of Rapid Differential Diagnosis of Classical Swine Fever Virus Antibody by E. Coli Expressed GFP Fusion Proteins
作者: 張天傑
劉昭君
王孟亮
關鍵字: 生物技術, 畜牧獸醫類
應用研究
摘要: Classical swine fever (CSF) causes one of the most severe diseases in pig worldwide. Infected pigs developed pyrexia and leukopenia. Mortality rates may reach 90%~100% when acute infect, and outbreak of classical swine fever can causes large economic consequences. In this project, we will construct prokaryotic plasmids that express CSFV core and N-terminal half of envelope protein E2 with enhanced green fluorescent protein (EGFP). The results of fluorescent microscope and luminescent spectrophotometer examinations of fusion proteins Core-EGFP and E2-EGFP, extracted from inclusion body, had been shown that these fusion proteins could absorb UV light and emit green fluorescence. In this study, a very rapid, sensitive and specific Dot-ELISA will be developed for immunological diagnosis in CSFV-infected swine. This immunological diagnosis method could be applied for differential diagnosis of CSFV marker vaccines and wildtype virus infection.
豬瘟病毒(classical swine fever virus, CSFV)會引起豬隻急性發熱性高度傳染性疾病-豬瘟.當豬隻感染豬瘟時會發生發燒、全身性出血、敗血等症狀.急性感染時,死亡率高達95%-100%,對養豬戶造成極大威脅與經濟損失.GFP體帶有由水母Aequorea victoria而來的綠色螢光素2 (green fluorecent protein;GFP)基因,於此E. coli的表現載體中,可經由T7 promotor及IPTG (Isopropryl-β-Thiogalactoside)的調控,所表現的蛋白質經長波長的子外線照射,可顯示出綠色螢光,而且此蛋白對細菌本身不會顯出毒性.本實驗可將豬瘟核蛋白或E2 蛋白嵌入此質體中並於E. coli中表現,所表現之融合蛋白產物可利用Dot-ELISA進行檢測其抗原性或以紫外線觀察其產生的螢光現象.此檢測方法將可利用表現蛋白作為豬瘟病毒快速鑑別診斷之新方法.
URI: http://hdl.handle.net/11455/52609
其他識別: 91農科-7.2.2-檢-B3(1)
文章連結: http://grbsearch.stpi.narl.org.tw/GRB/result.jsp?id=935184&plan_no=91%E8%BE%B2%E7%A7%91-7.2.2-%E6%AA%A2-B3%281%29&plan_year=91&projkey=PG9306-1658&target=plan&highStr=*&check=0&pnchDesc=%E4%BB%A5%E8%9E%A2%E5%85%89%E6%A8%99%E7%A4%BA%E8%9E%8D%E5%90%88%E8%9B%8B%E7%99%BD%E9%91%91%E5%88%A5%E8%A8%BA%E6%96%B7%E8%B1%AC%E7%98%9F%E6%8A%97%E9%AB%94%E4%B9%8B%E7%A0%94%E7%A9%B6
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