Transformation and replication in mouse cells of a bovine papillomavirus--pML2 plasmid vector that can be rescued in bacteria.
Transformation and replication in mouse cells of a bovine papillomavirus--pML2 plasmid vector that can be rescued in bacteria.
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牛乳头瘤病毒在小鼠细胞中的转化和复制——可在细菌中拯救的 pML2 质粒载体。
DOI：
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发表时间：
1982
影响因子：
11.1
通讯作者：
P. Howley
中科院分区：
文献类型：
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作者：
N. Sarver;J. Byrne;P. Howley
牛乳头瘤病毒（BPV-1）DNA在转化的小鼠细胞中作为稳定的多拷贝质粒复制的独特能力导致其用作真核克隆载体。该系统的一个局限性是当BPV-1 DNA或BPV-1 DNA的亚基因组转化片段（BPV 69 T）与pBR 322序列共价连接时，转化效率显著降低。构建了由BPV-1 DNA和pBR 322的缺失变体pML 2d组成的双宿主复制子，并显示其在体外对小鼠细胞的转化是高效的。该杂合分子在转化的小鼠细胞中作为稳定的、未整合的、多拷贝质粒复制。在细菌中拯救了驻留的BPV-1-pML 2d质粒DNA，并且回收的质粒结构与原始转化DNA相同，并且具有相同的转化效率。相反，当DNA与pML 2d共价连接时，BPV 69 T DNA的转化效率小于BPV-1 DNA的1/100。这些观察结果表明，虽然BPV-1（BPV 31 NT）DNA的非转化区是不是必不可少的转化，它有一个促进作用的转化过程。
The unique ability of bovine papillomavirus (BPV-1) DNA to replicate as a stable, multicopy plasmid in transformed mouse cells has led to its utilization as a eukaryotic cloning vector. One limitation of the system has been the marked reduction in transformation efficiency when BPV-1 DNA or the subgenomic transforming segment of BPV-1 DNA (BPV69T) is covalently linked to pBR322 sequences. A dual host replicon consisting of BPV-1 DNA and pML2d, a deletion variant of pBR322, was constructed and shown to be highly efficient for transformation of mouse cells in vitro. The hybrid molecule replicates as a stable, unintegrated, multicopy plasmid in transformed mouse cells. The resident BPV-1-pML2d plasmid DNA was rescued in bacteria and the recovered plasmids were shown to be identical in structure and to have the same transformation efficiency as the original transforming DNA. In contrast, the transforming efficiency of BPV69T DNA is less than 1/100th that of BPV-1 DNA when the DNA is left covalently linked to pML2d. These observations indicate that, although the nontransforming region of BPV-1 (BPV31NT) DNA is not essential for transformation, it has a facilitative role in the transformation process.