Robinson G W, Nicolet C M, Kalainov D, Friedberg E C
Proc Natl Acad Sci U S A. 1986 Mar;83(6):1842-6. doi: 10.1073/pnas.83.6.1842.
Plasmids containing various RAD-lacZ gene fusions were integrated into the chromosome of haploid yeast cells. These integrant strains were tested for expression of Escherichia coli beta-galactosidase after treatment with agents that damage DNA or interfere with normal DNA replication. We did not observe induction of single-copy RAD1-lacZ or RAD3-lacZ fusion genes under the experimental conditions used. However, exposure of cells containing an integrated RAD2-lacZ fusion gene to UV-radiation, gamma-radiation, 4-nitroquinoline 1-oxide, or nalidixic acid resulted in 4- to 6-fold enhanced expression of beta-galactosidase. Induction of the RAD2 gene after treatment of untransformed cells with 4-nitroquinoline 1-oxide was confirmed by direct examination of RAD2 mRNA. Lower levels of induction (approximately equal to 50%) were observed after treatment of cells with other chemicals. Induction of the RAD2-lacZ fusion gene was also observed in cells transformed with single-copy and multicopy autonomously replicating plasmids.
含有各种RAD-lacZ基因融合体的质粒被整合到单倍体酵母细胞的染色体中。在用损伤DNA或干扰正常DNA复制的试剂处理后,检测这些整合菌株中大肠杆菌β-半乳糖苷酶的表达。在所用的实验条件下,我们没有观察到单拷贝RAD1-lacZ或RAD3-lacZ融合基因的诱导。然而,含有整合的RAD2-lacZ融合基因的细胞暴露于紫外线、γ射线、4-硝基喹啉1-氧化物或萘啶酸后,β-半乳糖苷酶的表达增强了4至6倍。通过直接检测RAD2 mRNA证实了未转化细胞在用4-硝基喹啉1-氧化物处理后RAD2基因的诱导。用其他化学物质处理细胞后观察到较低水平的诱导(约50%)。在用单拷贝和多拷贝自主复制质粒转化的细胞中也观察到了RAD2-lacZ融合基因的诱导。