Tyndall C, Younghusband H B, Bellett A J
J Virol. 1978 Jan;25(1):1-10. doi: 10.1128/JVI.25.1.1-10.1978.
We have investigated the association of viral DNA with cell DNA in chicken embryo kidney (CEK) cells productively infected with chicken embryo lethal orphan (CELO) virus and in human (HEK) cells infected with mutants ts36 and ts125 of human adenovirus type 5 under permissive and restrictive conditions. Cell and viral DNA molecules were separated after CELO virus infection of CEK cells by alkaline sucrose gradient centrifugation, network formation, and CsCl density gradient centrifugation, methods that rely on different properties of the DNA. The cell DNA was then tested for viral sequences by DNA reannealing kinetics. Between 500 and 1,000 viral genome equivalents per cell were found at 36 h postinfection associated with cell DNA purified by each method. These values greatly exceeded the amount of free viral DNA found contaminating cell DNA prepared by the same methods from uninfected cells to which CELO virus DNA had been added. Quantitative agreement in the amounts of viral DNA found associated with cell DNA purified by these different methods suggests that CELO virus DNA is integrated into chick cell DNA during lytic infection. Similar experiments in HEK cells using mutants ts36 and ts125 of adenovirus type 5 at both restrictive and permissive temperatures showed that the same proportion of viral DNA is associated with cell DNA in the absence of viral DNA replication, and this suggests that the difference in the frequency with which cells are transformed by these mutants is not due to a difference in the frequency integration.
我们研究了在允许和限制条件下,鸡胚肾(CEK)细胞被鸡胚致死孤儿(CELO)病毒有效感染以及人胚肾(HEK)细胞被人腺病毒5型的ts36和ts125突变体感染时,病毒DNA与细胞DNA的关联。在CEK细胞被CELO病毒感染后,通过碱性蔗糖梯度离心、网状形成和CsCl密度梯度离心等方法分离细胞和病毒DNA分子,这些方法依赖于DNA的不同特性。然后通过DNA复性动力学检测细胞DNA中的病毒序列。在感染后36小时,每细胞发现500至1000个病毒基因组当量与通过每种方法纯化的细胞DNA相关联。这些值大大超过了从添加了CELO病毒DNA的未感染细胞中通过相同方法制备的污染细胞DNA中发现的游离病毒DNA的量。通过这些不同方法纯化的与细胞DNA相关联的病毒DNA量的定量一致性表明,CELO病毒DNA在裂解感染期间整合到鸡细胞DNA中。在HEK细胞中,使用腺病毒5型的ts36和ts125突变体在限制和允许温度下进行的类似实验表明,在没有病毒DNA复制的情况下,相同比例的病毒DNA与细胞DNA相关联,这表明这些突变体转化细胞的频率差异不是由于整合频率的差异。