Shih C C, Stoye J P, Coffin J M
Tufts University School of Medicine, Department of Molecular Biology and Microbiology, Boston, Massachusetts 02111.
Cell. 1988 May 20;53(4):531-7. doi: 10.1016/0092-8674(88)90569-7.
A central feature of retrovirus replication is integration of the provirus into host cell DNA, but the specificity of this step for cell target sequences has not been clarified. To investigate this issue, we developed a method for screening and comparing large numbers of unselected integration events. Using a replication-competent Rous sarcoma virus containing a bacterial suppressor tRNA gene as a selectable marker, we obtained collections of clones comprising integrated provirus together with host flanking sequences. Hybridization and sequence analysis of the flanking sequence reveals the presence of a number of strongly preferred integration targets. Within these targets, independent integration events occur at sites identical to the base.
逆转录病毒复制的一个核心特征是将前病毒整合到宿主细胞DNA中，但这一步骤对细胞靶序列的特异性尚未明确。为了研究这个问题，我们开发了一种筛选和比较大量未选择整合事件的方法。使用含有细菌抑制性tRNA基因作为选择标记的具有复制能力的劳氏肉瘤病毒，我们获得了包含整合前病毒以及宿主侧翼序列的克隆集合。侧翼序列的杂交和序列分析揭示了许多强烈偏好的整合靶点的存在。在这些靶点内，独立的整合事件发生在与碱基相同的位点。
