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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
關鍵字: 生物技術, 畜牧獸醫類
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.
其他識別: 91農科-7.2.2-檢-B3(1)
Appears in Collections:獸醫學系所



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