A shuttle vector plasmid for studying carcinogen-induced point mutations in mammalian cells.
A shuttle vector plasmid for studying carcinogen-induced point mutations in mammalian cells.
复制标题
用于研究哺乳动物细胞中致癌物诱导的点突变的穿梭载体质粒。
DOI：
10.1016/0378-1119(85)90222-7
复制
发表时间：
1985
期刊：
影响因子：
3.5
通讯作者：
Berman,ML
中科院分区：
文献类型：
--
作者：
Seidman,MM;Dixon,K;Razzaque,A;Zagursky,RJ;Berman,ML
我们已经构建了一个穿梭载体质粒，用于研究哺乳动物细胞中的诱变。该质粒在允许SV40病毒的细胞系以及大肠杆菌中复制，并携带可作为诱变标记的细菌抑制tRNA基因（supF）。该质粒在非洲绿色猴肾CV1细胞中与SV 40病毒一样有效地复制，表明在pBR 322及其衍生物中通常发现的所有痕量抑制序列已被去除。质粒的设计和诱变靶基因的小尺寸降低了恢复自发缺失突变的概率，所述自发缺失突变已被证明在哺乳动物细胞中传代期间以高频率发生。在CV1细胞中传代后回收的自发突变质粒的频率显著低于先前描述的其它载体。当质粒DNA在CV1细胞中传代前用UV辐射处理时，观察到突变体的频率约为自发背景的20倍。
We have constructed a shuttle vector plasmid for studying mutagenesis in mammalian cells. The plasmid replicates in cell lines permissive for SV40 virus as well as in the bacteriumEscherichia coliand carries a bacterial suppressor tRNA gene (supF) that can serve as a mutagenesis marker. The plasmid replicates as efficiently as SV40 virus in African Green Monkey kidney CV1 cells, indicating that all traces of the inhibitory sequences normally found in pBR322 and its derivatives have been removed. The design of the plasmid and the small size of the mutagenesis target gene decrease the probability of recovering spontaneous deletion mutations that have been shown to occur at high frequency during passage in mammalian cells. The frequency of spontaneous-mutant plasmids recovered after passage in CV1 cells is substantially lower than with other vectors described previously. When the plasmid DNA is treated with UV radiation before passage in CV1 cells, mutants are observed at a frequency about 20-fold above the spontaneous background.