https://scholars.lib.ntu.edu.tw/handle/123456789/136139
標題: | 魚類結病毒與魚類反轉錄病毒共感染時之干擾研究(一) | 作者: | 齊肖琪 | 關鍵字: | 石斑魚神經壞死症病毒;鱧魚反轉錄病毒;共感染;石斑魚鰭細胞株;grouper nervous necrosis virus (GNNV);snakehead retrovirus (SnRV);coinfection;grouper fin cell line (GF-1 cell line) | 公開日期: | 2004 | 出版社: | 臺北市:國立臺灣大學生命科學系 | 摘要: | 本計劃在探討石斑魚神經壞死症病毒(Grouper Nervous Necrosis Virus, GNNV) 和鱧魚反轉錄病毒(Snakehead Retrovirus, SnRV) 共感染同一寄主細胞株時會發生的 干擾現象。將SnRV 感染石斑魚鰭細胞株GF-1,使之成為有SnRV 持續性感染的 SGF-1 細胞。在SGF-1 細胞核酸萃取物中可測到 SnRV 的cDNA,細胞上清液中 測到SnRV 的RNA,又用SGF-1 細胞上清液感染新的GF-1 細胞,感染後的細胞 核酸萃取物中可再度測到SnRV 的cDNA,證明SGF-1 細胞上清液中有感染活性的 SnRV 病毒顆粒,因此證明SnRV 在SGF-1 細胞中可以完成生活史。將GNNV 以 相同感染劑量(Multiplicity of infection, MOI),同步感染相同細胞數目的GF-1 及 SGF-1 兩組細胞,並觀察兩組細胞中病毒核酸與力價的日變化,結果顯示 (1) SnRV 的反轉錄酶 (Reverse Transcriptase, RT) 會完整地把GNNV 的兩條單股遺傳RNA 序列反轉錄為cDNA,使GNNV 生活史中產生了原來沒有的新階段;(2) GNNV 的 cDNA 於感染後的第三天開始出現,且隨感染時間增加而增加;(3) GNNV RNA 的 產量在兩組細胞系統中沒有差異;(4) SnRV RNA 的量在SGF-1 細胞中會隨著感染 GNNV 天數的增加而遞減;(5) SGF-1 細胞感染GNNV 第三天到第五天所收的上清 液,在GF-1 和SGF-1 兩個系統中所定出來的力價有明顯差距,第四天的上清液在 GF-1 細胞中定出來的力價比在SGF-1 細胞定出來的力價高出104 倍,推測力價的 差距與GF-1 細胞上有兩種病毒的受器可用,而SGF-1 細胞因SnRV 的持續性感染 造成SnRV 受器的關閉,只有GNNV 受器可用有關連。 The aim of this study is to investigate the interference of grouper nervous necrosis virus (GNNV) and snakehead retrovirus (SnRV) during the co-infection of the same host cell line. A SnRV-persistent infected cell line SGF-1 was induced by inoculating the SnRV into GF-1 cell line. The cDNA of SnRV was detected in the nucleic acid extract of SGF-1 cells, and the genomic RNA of SnRV was also detected in the culture supernatant of SGF-1 cells. Moreover, the supernatant of SGF-1 cells was used for the infection of GF-1 cells, and SnRV cDNA was once again detected in infected GF-1 cells indicating that there were infectious particles of SnRV in the supernatant of SGF-1 cells, and the life cycle of SnRV could be completed in the SGF-1 cells. Synchronous infection of GNNV in GF-1 cells and SGF-1 cells was done for the observation of daily changes of the nucleic acids and the titers of the viruses. The results revealed that (1) the SnRV reverse transcriptase (RT) could completely reverse-transcribed GNNV single-stranded RNA1 and RNA2 into cDNA during the co-infection in SGF-1 cells, and created a new cDNA stage in the life cycle of GNNV; (2) the cDNA appeared in SGF-1 cells since the third day after GNNV infection, and the amount of the cDNA increased as the days increased; (3) the amount of GNNV RNA in either infected GF-1 cells or SGF-1 cells were very similay; (4) the SnRV RNA decreased as the infection days increased; (5) the titers of the supernatants of GNNV-infected SGF-1 cells during the third day to the fifth day determined in GF-1 cells were very different from the titers determined in SGF-1 cells, and the titer of the fourth day determined in GF-1 cells was 104 times higher than that determined in SGF-1 cells. This phenomenon could be related with that the receptors of GNNV and SnRV were available on GF-1 cells, while the receptors of SnRV were closed on SGF-1 cells owing to the SnRV-persistent infection, and only GNNV receptors were available on SGF-1 cells. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/10282 | 其他識別: | 922311B002105 | Rights: | 國立臺灣大學生命科學系 |
顯示於: | 生命科學系 |
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