AAV REP PROTEINS AND TARGETED INTEGRATION

腺相关病毒(AAV)DNA靶向整合入人 19号染色体通过非同源重组发生。出席会议的人 AAV非结构蛋白(Rep)基因与特异性相关 前向整合。因此，我们调查了 靶向整合中的Rep蛋白。为分析代表提供便利 蛋白质功能、生物活性、重组Rep蛋白 在大肠杆菌中生产。Rep与线性双链DNA的结合是一种新的 重新定义绑定要求的Rep的标识属性 Rep蛋白。AAV DNA 19号染色体整合位点是一种特殊的 用于Rep结合底物，建立Rep可以作为 有针对性的整合。体外复制实验结果 证明了19号染色体的亚片段可以作为Rep特异性的 复制的来源。因此，我们认为在AAV感染的细胞中，Rep 促进从集成位置进行有限复制。亲和力 病毒或宿主底物的Rep蛋白可以允许Rep 与聚合酶复合体的亚基交换导致分子间 DNA重组。

Targeted integration of adeno-associated viral (AAV) DNA into human chromosome 19 occurs by nonhomologous recombination. The presence of the nonstructural protein (Rep) genes of AAV correlated with the specificity of proviral integration. Therefore, we investigated the involvement of Rep proteins in targeted integration. To facilitate the analysis of Rep protein functions, biologically active, recombinant Rep proteins were produced in E. coli. Binding of Rep to linear duplex DNA is a newly identified property of Rep which redefines the binding requirements for Rep proteins. The AAV DNA chromosome 19 integration locus is a specific substrate for Rep binding, establishing that Rep may act as an agent for targeted integration. The results of in vitro replication experiments demonstrated that a chromosome 19 subfragment may act as a Rep specific origin of replication. Thus we believe that in AAV infected cells, Rep promotes limited replication from the integration locus. The affinity of Rep proteins for either the viral or host substrate may permit Rep subunit exchange with the polymerase complex resulting in intermolecular DNA recombination.