Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/93009
標題: The Characterization of Chicken Infectious Anemia Virus-like Particles Expressed by A Recombinant Fowlpox Virus
以重組禽痘病毒表現雞傳染性貧血類病毒顆粒特性之研究
作者: Yi-Chia Lo
羅翊嘉
關鍵字: 雞傳染性貧血病毒;重組禽痘病毒;chicken infectious anemia virus;recombinant fowlpox virus
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摘要: 
雞傳染性貧血 (chicken infectious anemia; CIA)主要由雞傳染性貧血病毒(chicken infectious anemia virus; CIAV)造成,此病毒分布全球影響所有國家且尤其對於肉雞產業及提供無特定病原蛋之生產者造成經濟威脅,雖然目前市面上有活毒疫苗用於免疫但推薦施打日齡皆超過6週齡,對於幼齡雞隻無法提供完全的保護。因此,本研究目的為構築一質體嵌入目標基因後建立重組禽痘病毒(fowlpox virus; FPV),藉由同源重組共同表現CIAV VP1及VP2基因,其中,CIAV基因來源為收集自臨床病例,經過選殖及定序決定一株台灣地區野外高致病力分離株。構築一包含thymidine kinase (TK)、green fluorescent protein (GFP)、CIAV VP1及VP2之重組質體,TK基因藉特異限制切位分成兩部分並嵌入pGEX6P-1質體之多重選殖位上,TK基因的上下游位於GFP、CIAV VP1及VP2三種蛋白基因的兩側並嵌入載體以產生pGEX6P-1/TK/GFP/CIAV VP1/VP2重組質體。接著,為產生重組禽痘病毒,將禽痘病毒作為父母代病毒感染於DF1雞纖維母細胞,同時和重組質體進行轉染。父母代禽痘病毒和重組質體成功進行同源重組後可於螢光顯微鏡下看到綠色螢光表現,觀察到第一代重組禽痘毒之產生後,進行間接免疫螢光試驗以確定CIAV蛋白的表現。

Chicken infectious anemia virus (CIAV) is the causative agent of chicken infectious anemia (CIA). CIA distributes globally, affecting all countries and presenting economic threat especially to the broiler industry and the producers of specific-pathogen (SPF) eggs. Though live vaccines are now in use, the recommended age is over 6 weeks of age and can't offer complete protection to young chickens. Thus, the purpose of this study was to construct the plasmid which inserted target genes and developed a recombinant fowlpox virus (rFPV) co-expressing VP1 and VP2 genes of CIAV by homologous recombination. The strain of CIAV was sampled from clinical cases and helped determine a high pathogenicity strain after cloning and sequencing. A recombinant plasmid was constructed with TK, GFP, VP1 and VP2 genes of CIAV. TK gene was separated to 2 segments by specific restriction sites and was inserted into the on multiple cloning sites of the pGEX6P-1 vector. The separated TK gene segments flanked GFP gene, VP1 and VP2 genes of CIAV were inserted in the pGEX6P-1 vector to generate a pGEX6P-1/TK/GFP/CIAV VP1/VP2 recombinant plasmid. Next, to generate rFPV, DF1 cells were infected by FPV as parental FPV and then transfected with recombinant plasmid at the same time. After homologous recombination with parental FPV, green fluorescence could be seen by fluorescence microscope when recombination occurred. After observing the first generation of recombinant fowlpox virus, indirect immunofluorescence assay (IFA) tests were performed to confirm the expression of CIAV proteins.
URI: http://hdl.handle.net/11455/93009
其他識別: U0005-2707201518475400
Rights: 不同意授權瀏覽/列印電子全文服務
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