Lebkowski J S, DuBridge R B, Antell E A, Greisen K S, Calos M P
Mol Cell Biol. 1984 Oct;4(10):1951-60. doi: 10.1128/mcb.4.10.1951-1960.1984.
Papovavirus-based shuttle vectors containing the bacterial lacI gene were used to show that a mutation frequency in the range of 1% occurs in lacI when such vectors are transfected into COS7 and CV-1 simian cells, NIH 3T3, 3T6, L, and C127 mouse cells, and human 293 and HeLa cells. This frequency is approximately four orders of magnitude higher than the spontaneous mutation frequency in either mammalian or bacterial cells. The mutations are predominantly base substitutions and deletions and also include insertions from the mammalian genome. Time course experiments argue that mutagenesis occurs soon after arrival of the DNA into the nucleus. However, replication of the vector is not required since mutations occur even when the vector lacks all viral sequences. The high mutation frequency appears to be the characteristic outcome of transfection of DNA into mammalian cells.
含有细菌lacI基因的基于乳头瘤病毒的穿梭载体被用于证明,当将此类载体转染到COS7和CV-1猴细胞、NIH 3T3、3T6、L和C127小鼠细胞以及人293和HeLa细胞中时,lacI基因的突变频率在1%的范围内。该频率比哺乳动物或细菌细胞中的自发突变频率大约高四个数量级。这些突变主要是碱基替换和缺失,也包括来自哺乳动物基因组的插入。时间进程实验表明,诱变在DNA进入细胞核后不久就会发生。然而,载体的复制并非必需,因为即使载体缺乏所有病毒序列,突变也会发生。高突变频率似乎是将DNA转染到哺乳动物细胞中的典型结果。