Kitamura Y, Yoshikura H, Kobayashi I
Department of Bacteriology, Faculty of Medicine, University of Tokyo, Japan.
Mol Gen Genet. 1990 Jul;222(2-3):185-91. doi: 10.1007/BF00633816.
Bovine papillomavirus (BPV) shuttle vectors replicate as a circular plasmid in mouse cell nuclei without impairing host cell viability. We used these vectors to analyze homologous recombination in mammalian cells. When several BPV-based plasmids carrying direct repeats were introduced into C127 cells, we detected many recombinant plasmid molecules that have lost the sequence between the repeats. Many recombinant type molecules as well as parental type molecules were detected in all the cell clones isolated for analysis. Sequencing after rescue of the plasmid in Escherichia coli showed that most of the recombinants were from accurate homologous recombination. When the repeats on the plasmid were in inverted orientation, no crossing-over type products were detected. We discuss possible mechanisms that explain these features.
牛乳头瘤病毒（BPV）穿梭载体在小鼠细胞核中作为环状质粒进行复制，而不会损害宿主细胞的活力。我们使用这些载体来分析哺乳动物细胞中的同源重组。当将几个携带同向重复序列的基于BPV的质粒导入C127细胞时，我们检测到许多失去了重复序列之间序列的重组质粒分子。在所有分离用于分析的细胞克隆中都检测到了许多重组型分子以及亲本型分子。在大肠杆菌中拯救质粒后进行测序表明，大多数重组体来自精确的同源重组。当质粒上的重复序列呈反向排列时，未检测到交叉型产物。我们讨论了解释这些特征的可能机制。
