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題 名 | Development of a Penaeus Monodon-Type Baculovirus (MBV) Probe by Polymerase Chain Reaction and in Situ Hybridization=應用聚合酶連鎖反應和原位雜交反應建立草蝦桿狀病毒檢驗探針 |
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作 者 | 盧秀琴; 陳秀男; | 書刊名 | 臺北師院學報 |
卷 期 | 8 1995.06[民84.06] |
頁 次 | 頁377-391 |
分類號 | 387.13 |
關鍵詞 | 聚合酶; 連鎖反應; 原位雜交反應; 草蝦; 桿狀病毒; 檢驗; Penaeus Monodon-Type Baculovirus; MBV; |
語 文 | 英文(English) |
中文摘要 | 利用蔗糖梯度離心技術,將草蝦桿狀病毒(Penaeus monodon-type Baculovirus; MBV)自感染MBV的草蝦苗(PL10-20)肝胰腺中純化出來,萃取MBV DNA後, 參考MBV的多角體基因(Polyhedrosis gene)設計引子(Primers),對MBV DNA進行聚合 ?連鎖反應(ploymerase chain reaction; PCR),得到一段650個鹼基對(base pair)PCR產 物;用digoxigenin-dUTP來標識此段PCR產物,藉由點墨雜交反應,發現此段標識的DNA, 對於MBV具有專一性和很高的靈敏度,可以當作檢驗探針。利用此檢驗探針當作檢驗工具, 對草蝦組織做原位雜交反應(in situ hybridization),發現MBV感染草蝦的肝胰腺和中腸 組織。在蝦苗的眼幼蟲期(zoea)即可以檢驗到有MBV的感染,眼幼蟲期之前則偵測不到。 |
英文摘要 | Penaeus monodon-type baculovirus (MBV) was isolated and purified from the hepatopancreata of MBV-infected Penaeus monodon Fabricius. MBV DNA was extracted and used as a template in polymerase chain reaction (PCR). The primers were chosen from the polyhedron gene of MBV. One DNA fragment was amplified after PCR. The DNA fragment was subsequently used as a probe in a dot blot analysis; the result showed that the probe was able to hybridize to the DNA extracted from the purified MBV and from the MBV-infected P. monodon, but not from the MBV-uninfected P. monodon. The probe, labeled with digoxigenin, was used for in situ hybridization. MBV-infected tissue sections were microscopically examined for a blue-purple precipitate indicative of a positive reaction for MBV. MBV-positive cells were located only in the hepatopancreatic tubule epithelium and in the midgut epithelium. |
本系統中英文摘要資訊取自各篇刊載內容。