J亚型禽白血病病毒的拯救与鉴定

摘要

为鉴定含J亚群禽白血病病毒(ALV-J)全长cDNA分子克隆(pBlALV)的感染性,首先将pBlALV转染CEF细胞,拯救出病毒(rALV),然后连续传代3次,对拯救病毒进行增殖培养.分别利用RT-PCR、western blot、IFA等方法,对拯救病毒进行鉴定,并应用实时荧光定量PCR方法对拯救病毒的复制动力学进行了测定.研究结果证明,本研究成功拯救出了具有感染性的rALV-J,为研究禽白血病的致病机理和探讨新的防制措施等提供了良好的ALV的反向遗传操作技术平台.%To confirm the infectivity of the fiill-length cDNA clone (pBlALV) of subgroup J avian leucosis virus (ALV-J) HPRS103 strain, pBlALV was transfected into chicken embryo fibroblast (CEF) cells and the ALV was rescued. The rescued ALV (rALV-J) was identified by RT-PCR, western blot and indirect immunofiuorescence assay, respectively. Besides, the real-time PCR was applied to evaluate the replication dynamic of the rescued virus. The results showed that the rALV-J was able to well replicate in CEF. This study provides an useful platform for investigation of the pathogenesis and molecular biology of ALV-J.