請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75478
標題: | 日本鰻所分離出兩段核醣核酸病毒之特性研究 Characteristics of A Birnavirus Isolated from Japanese Eel (Anguilla japonica) |
作者: | Jih-Yau Chang 張繼堯 |
出版年 : | 1983 |
學位: | 碩士 |
摘要: | 由具有鰭部充血、體表潰爛、肝臟腫大壞死和體軀凹凸等病徵之日本鰻的肝臟、脾臟和腎臟,都可分離出病毒。利用細胞培養病毒,發現其在CHSE-214細胞中增殖最好。代謝抑制物FUDR和Actinomycin D試驗,結果顯示病毒之遺傳物質為核醣核酸。電子顯微鏡觀察下,病毒存在細胞質中靠近內質網處。負染色之純化病毒,顯示本病毒為不具外套膜,且祇含單層外套之二十面體,直徑約為59.8nm。在氯化銫中的浮力密度為1.33g/ml•SDS-polyacrylamide膠體電泳分析純化病毒之蛋白質和核醣核酸,顯示病毒顆粒含有分子量分別為90x10 3(VP 90),46x10 3(VP 46)和32x10 3(VP 32) daltons之蛋白質帶,以14C-uridine標示之病毒經電泳分析,由自動放射攝影中顯示出兩段核醣核酸。在低能量紫外線照射下,病毒之殘存率呈肩形曲線遞減,顯示其核醣核酸為雙股。中和試驗結果顯示其血清型和EVE(Eel Virus,European)密切相關,由以上結果證實本病毒屬於Birnaviridae病毒中的EVE病毒.但是進一步比較14C-amino acid標示之此分離病毒和Sano分離的EVE病毒的蛋白質,發現其在電泳膠體中的泳動距離和含量百分比略有不同。而利用紫外線照射和virazole處理抑制病毒增殖之實驗,發現在15,000 J.m-2劑量的紫外線下,可有效降低病毒感染力10 7TCID50/ml.1μg/ml劑量的virazole即可抑制病毒感染力10 4.5 TCID 50/ml。經由14C-uridine和14C-amino acid標示研究結果,virazole是抑制在病毒核醣核酸的合成。 The virus was isolated from the liver, kidney and sp1 of diseased Japanese eels. Affected eels showed ulcer lesio over the body, congestion of fins, hypertrophy and necrosis of the liver, atrophic muscle and deformed trunk. Among the multiplication test of virus in various cell lines, we found that CHSE-214 cell line was the best one for viral multiplication. The metabolic inhibition test indicated that the viral nucleic acid was RNA. Electron microscopic observation revealed that the virus was presented in the cytoplasm near endoplasmic reticulum. It was unenveloped icosahedral, approximately 59.8 nm in diameter, and only had single layer capsid. The buoyant desity of the isolated virus in CsCl was 1.33 g/ml. By using polyacrylamide gel electrophoresis, three protein bands could be observed from the isolated virus, there were VP90 (M.W. 90,000 daltons), 46 (M.W. 46,000 daltons) and VP32 (M.W. 32,000 daltons). Autoradiograph showed that the virus possessed two segments of RNA genome. Under TJV light irradiation at low fluences, survial curve of virus displaied minor shoulder which revealed that the viral RNA was double stranded. The neutralization test indicated that the isolated virus had similar serotype with EVE (Eel Virus, European), and could be classified into EVE group of Binaviridae. But there were some differences in protein mobility and composition percentages between the isolated virus and EVE. The viral multiplication was inhibited by UV irradition and virazole treatment. Viral infectivity was reduced by 10 7 TCID50/ml under 15,000 J. m-2 tensity of UV irradiation, and was inhibited by 10 4.5TCID50/ml when 1 μg/ml of virazole was treated. Virazole inhibited the viral ribonucleic acid synthesis which was confirmed by using 14C-uridine and acid tracing. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75478 |
全文授權: | 未授權 |
顯示於系所單位: | 動物學研究所 |
文件中的檔案:
沒有與此文件相關的檔案。
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。