DETECTION AND ANALYSIS OF UV-INDUCED MUTATIONS IN MAMMALIAN-CELL DNA USING A LAMBDA-PHAGE SHUTTLE VECTOR
DETECTION AND ANALYSIS OF UV-INDUCED MUTATIONS IN MAMMALIAN-CELL DNA USING A LAMBDA-PHAGE SHUTTLE VECTOR
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DOI：
10.1073/pnas.83.4.1041
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发表时间：
1986-02-01
影响因子：
11.1
通讯作者：
SUMMERS, WC
中科院分区：
文献类型：
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作者：
GLAZER, PM;SARKAR, SN;SUMMERS, WC
为了研究哺乳动物细胞的诱变作用，建立了多拷贝的小鼠L细胞系。含有大肠杆菌supF基因的噬菌体载体，作为诱变的目标。用.lambda从高分子量小鼠细胞DNA中拯救活噬菌体。体外包装提取物是有效的(每个拷贝的细胞DNA每微克5个噬菌体)，并且产生的突变噬菌体的背景可以忽略不计(54,605个中的0个)。从254 nm紫外光(12J/m2)照射的小鼠细胞中，共救出78510个噬菌体，其中8个发现有突变的supF基因。对突变体的DNA序列分析表明，哺乳动物细胞中紫外线诱变的主要位置在嘧啶-胞嘧啶(Y-C)序列上，该位置最常见的突变是C.fwdarw。T转换。
In order to study mutagenesis in mammalian cells, stable mouse L-cell lines were established with multiple copies of a .lambda. phage vector that contains the supF gene of Escherichia coli as a target for mutagenesis. Rescue of viable phage from high molecular weight mouse cell DNA using .lambda. in vitro packaging extracts was efficient (5 phage per .mu.g of cell DNA per copy) and yielded a negligible background of mutant phage (0 out of 54,605). From mouse cells exposed to 254-nm ultraviolet light (12 J/m2), 78,510 phage were rescued, of which 8 were found to have mutant supF genes. DNA sequence analysis of the mutants suggests that the primary site of UV mutagenesis in mammalian cells is at pyrimidine-cytosine (Y-C) sequences, and that the most frequent mutation at this site is a C .fwdarw. T transition.