SITE-SPECIFIC INTEGRATION BY ADENOASSOCIATED VIRUS
SITE-SPECIFIC INTEGRATION BY ADENOASSOCIATED VIRUS
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DOI：
10.1073/pnas.87.6.2211
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发表时间：
1990-03-01
影响因子：
11.1
通讯作者：
BERNS, KI
中科院分区：
文献类型：
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作者：
KOTIN, RM;SINISCALCO, M;BERNS, KI
腺相关病毒(AAV)基因组整合拷贝两侧的细胞序列从潜伏感染的克隆人细胞系中分离出来，并用于探测来自另外21个独立获得的潜伏感染AAV克隆的人类细胞的基因组斑点。在未感染的人类细胞系和原生人类组织的基因组印迹中，每个侧翼序列探针都杂交到独特的条带，但在22个潜伏感染克隆中的15个中，侧翼序列不仅与原始片段杂交，还与总共36个额外物种杂交。AAV探针还与其中22条新条带杂交，代表了15个阳性克隆中的11个，但从未与未感染细胞DNA的片段特征杂交。根据这些数据，我们得出结论，AAV基因组优先整合到细胞基因组的特定区域。我们已经通过体细胞杂交作图确定了该整合位点是19号染色体所特有的，并且该序列已经从未感染的人类DNA中分离出来。
Cellular sequences flanking integrated copies of the adeno-associated virus (AAV) genome were isolated from a latently infected clonal human cell line and used to probe genomic blots derived from an additional 21 independently derived clones of human cells latently infected with AAV. In genomic blots of uninfected human cell lines and of primary human tissue, each flanking-sequence probe hybridized to unique bands, but in 15 of the 22 latently infected clones the flanking sequences hybridized not only to the original fragments but also to a total of 36 additional species. AAV probes also hybridized to 22 of these new bands, representing 11 of the 15 positive clones, but never to the fragment characteristic of uninfected cell DNA. From these data we conclude that the AAV genome preferentially integrates into a specific region of the cellular genome. We have determined that the integration site is unique to chromosome 19 by somatic cell hybrid mapping, and this sequence has been isolated from uninfected human DNA.