ADENO-ASSOCIATED VIRUS IN VITRO INTEGRATION AND REPLICATION

腺相关病毒(AAV)DNA优先整合到 人类19号染色体上的特定位点通过非同源重组。 病毒非结构蛋白Rep与蛋白质的相互作用 19号染色体衍生的DNA和AAV内部末端重复序列(ITR) 提示病毒蛋白参与了靶向整合。我们 认为有针对性的整合是有限的DNA复制造成的 由Rep与共识结合序列存在的结合启动 整合位点和病毒ITR。我们假设一个 涉及细胞DNA合成受限的动力学机制 Rep亚基之间的序列和蛋白质-蛋白质相互作用 蛋白质和细胞DNA聚合酶复合体产生 病毒-细胞重组连接。广义和广义 我们已经确定的代表函数与该模型一致。 潜伏感染细胞系的基因组DNA分析是支持的 以及细胞序列的复制和重排 都在现场。此外，到目前为止，每一个AAV前病毒的特征都是 相对于Rep结合呈不对称分布 元素。

Adeno-associated virus (AAV) DNA integrates preferentially into a specific locus on human chromosome 19 by non-homologous recombination. The interactions of the non-structural viral protein, Rep, with chromosome 19-derived DNA and AAV internal terminal repeats (ITR) suggests involvement of the viral protein in targeted integration. We propose that targeted integration results from limited DNA replication initiated by binding of Rep to a consensus binding sequencing present in both the integration locus and the viral ITR. We postulate that a dynamic mechanism involving both limited DNA synthesis of the cellular sequences and protein- protein interactions between subunits of Rep protein and a cellular DNA polymerase complex generates the viral-cellular recombination junctions. The generalized and extended Rep functions we have determined are consistent with this model. Analysis of genomic DNA of latently infected cell lines are supportive as well in that duplications and rearrangements of cellular sequences are present. Furthermore, every AAV provirus characterized to date was present asymmetrically distributed with respect to the Rep-binding element.