【摘要】 鸡传染性贫血病毒(CIAV)感染鸡引起免疫抑制性疾病，严重危害养禽业。为了构建CIAV Cux-1株的感染性克隆，本试验通过PCR扩增CIAV全基因组序列，与pcDNA3.1载体同源重组，得到重组质粒pcDNA3.1-Cux-1。进一步通过对pcDNA3.1-Cux-1酶切、环化得到CIAV Cux-1环状DNA。将环状DNA电转染导入MSB1细胞并连续传代，用Western blot和间接免疫荧光进行重组病毒的鉴定。结果显示，本试验成功拯救出CIAV重组病毒rCux-1 (recombinant Cux-1),重组病毒rCux-1携带有特定的遗传标记，并且与亲本株复制能力和蛋白表达水平基本一致。本试验成功构建的CIAV Cux-1株感染性克隆为进一步研究CIAV的致病机理和研制新型疫苗提供了试验基础。更多还原

【Abstract】 Chicken infectious anemia virus(CIAV) causes immunosuppressive diseases in chickens and seriously threatens the poultry industry. In order to construct an infectious clone of CIAV Cux-1 strain, whole genome sequence of the virus was amplified by PCR and homologously recombined with pcDNA3.1 vector to obtain recombinant plasmid pcDNA3.1-Cux-1. CIAV Cux-1 cyclic DNA was obtained by enzymatic cleavage of pcDNA3.1-Cux-1 and further cyclization. Cyclic DNA was introduced into MSB1 cells by electrotransfection. Western blot and indirect immunofluorescence assay indicated that the recombinant virus rCux-1 was successfully rescued through successive passages. The recombinant virus rCux-1 carried specific genetic markers and was essentially similar to the parental strain in replication and protein expression level. Thus, the infectious clone of CIAV Cux-1 strain constructed in this study offers a valuable tool for further study of CIAV pathogenesis and development of novel vaccines.更多还原