MOUSE CELLS TRANSFORMED BY BOVINE PAPILLOMAVIRUS CONTAIN ONLY EXTRACHROMOSOMAL VIRAL-DNA SEQUENCES
MOUSE CELLS TRANSFORMED BY BOVINE PAPILLOMAVIRUS CONTAIN ONLY EXTRACHROMOSOMAL VIRAL-DNA SEQUENCES
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DOI：
10.1073/pnas.78.5.2727
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
HOWLEY, PM
中科院分区：
文献类型：
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作者：
LAW, MF;LOWY, DR;HOWLEY, PM
通过重关联动力学和印迹杂交检查用 1 型牛乳头瘤病毒 (BPV-1) 病毒粒子、全长线性 BPV-1 DNA 或 BPV-1 的确定的转化亚基因组 DNA 片段转化的小鼠 C127 细胞中的病毒 DNA 序列。在所有情况下，转化细胞均含有多个 BPV-1 DNA 拷贝，仅以超螺旋或带切口的环状染色体外分子或缓慢迁移的环状病毒 DNA 分子复合物的形式存在。在用全长线性 BPV-1 DNA 转染的细胞建立的转化细胞系中，输入 DNA 发生再环化，这在某些情况下导致 DNA 线性化中使用的限制性位点丢失。在用定义的亚基因组片段建立的转化细胞系中，DNA发生环化，并伴随着新序列的获得或BPV-1序列的复制和重排。与其他经过充分研究的病毒转化系统相比，在足够敏感以检测 0.1-0.2 病毒基因组当量的条件下，未检测到 BPV-1 基因组与宿主染色体的整合。转化的维持可以由非整合的病毒DNA介导。
The viral DNA sequences in mouse C127 cells transformed by bovine papillomavirus type 1 (BPV-1) virions, by full-length linear BPV-1 DNA, or by a defined transforming subgenomic DNA segment of BPV-1 were examined by reassociation kinetics and blot hybridization. In all cases, the transformed cells contained multiple copies of BPV-1 DNA, present exclusively as supercoiled or nicked circular extrachromosomal molecules or as a slowly migrating complex of circular viral DNA molecules. In the transformed cell lines established from cells transfected with full-length linear BPV-1 DNA, there was recircularization of the input DNA which in some cases resulted in the loss of the restriction site used in the linearization of the DNA. In the transformed cell lines established with the defined subgenomic segment there was circularization of the DNA accompanied by the acquisition of new sequences or duplication and rearrangement of the BPV-1 sequences. In contrast to other well-studied virus transformation systems, no integration of the BPV-1 genome into the host chromosome was detected under conditions sensitive enough to detect 0.1-0.2 viral genome equivalent. Maintenance of transformation may be mediated by nonintegrated viral DNA.