The lacI shuttle: rapid analysis of the mutagenic specificity of ultraviolet light in human cells.
The lacI shuttle: rapid analysis of the mutagenic specificity of ultraviolet light in human cells.
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DOI：
10.1073/pnas.82.24.8606
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发表时间：
1985-12
影响因子：
11.1
通讯作者：
J. Lebkowski;S. Clancy;J. Miller;M. Calos
中科院分区：
文献类型：
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作者：
J. Lebkowski;S. Clancy;J. Miller;M. Calos
已经设计出一种系统，可以很容易地在DNA序列水平上分析作用于人类细胞的诱变剂的影响。携带在穿梭载体上的细菌基因LacI通过转基因进入人类组织培养细胞，并允许在细胞核内复制。在转基因后24到48小时，细胞暴露在诱变剂中。经过1-2天的进一步复制后，提纯载体DNA，并将其转回大肠杆菌进行评分和LacI突变分析。用这种方法推导出了53个紫外光诱导突变的核苷酸序列变化。大多数突变是过渡性的，发生在嘧啶-嘧啶序列上。在大肠杆菌中观察到的诱变特异性与紫外光非常相似，这表明人类和细菌细胞对紫外光的损伤反应相似。以这种方式使用LACI航天飞机应该可以确定人类细胞中广泛的诱变剂和致癌物的诱变特异性。
A system has been devised that allows the effect of mutagens acting in human cells to be readily analyzed at the DNA sequence level. The bacterial gene lacI, carried on a shuttle vector, is introduced into human tissue culture cells by transfection and allowed to replicate in the cell nucleus. Twenty-four to 48 hr after transfection, the cells are exposed to a mutagen. After 1-2 days of further replication, vector DNA is purified and transfected back into Escherichia coli for scoring and analysis of mutations in lacI. The nucleotide sequence changes for 53 UV light-induced mutations have been deduced in this way. Most of the mutations are transitions and occur at pyrimidine-pyrimidine sequences. The mutagenic specificity observed closely resembles that of UV light in E. coli, suggesting that human and bacterial cells respond similarly to damage from UV light. Use of the lacI shuttle in this way should permit determination of the mutagenic specificity of a wide range of mutagens and carcinogens in human cells.