4'-羟甲基-4,5',8-三甲基补骨脂素对M13 mp10噬菌体DNA中乳糖启动子区域的诱变作用
Mutagenesis of the lac promoter region in M13 mp10 phage DNA by 4'-hydroxymethyl-4,5',8-trimethylpsoralen.
作者信息
Piette J, Decuyper-Debergh D, Gamper H
出版信息
Proc Natl Acad Sci U S A. 1985 Nov;82(21):7355-9. doi: 10.1073/pnas.82.21.7355.
Abstract
Double-stranded M13 phage DNA (M13 mp10 replicative form) was photoreacted with 4'-hydroxymethyl-4,5',8-trimethylpsoralen, using light of wavelength greater than 320 nm or greater than 390 nm to generate predominantly crosslinks or monoadducts, respectively. The damaged DNAs were scored for inactivation and mutagenesis after transfection into Escherichia coli. The appearance of light-blue or colorless plaques on indicator medium showed that mutation had occurred in the lac insert of the viral DNA. A comparison of the consequences of the two phototreatments with psoralen supports the idea that crosslinks are both more lethal and more mutagenic than monoadducts. Numerous mutant clones partially or totally deficient in beta-galactosidase were plaque-purified and amplified. The viral DNA of each clone was sequenced by the dideoxy chain-terminating procedure. All of the observed base-pair changes were mapped to the lac promoter region and consisted of 3 transition, 14 transversion, and 6 single base-pair frame-shift mutations. The predominant mutation was a T.A----G.C transversion.
摘要
双链M13噬菌体DNA（M13 mp10复制型）与4'-羟甲基-4,5',8-三甲基补骨脂素进行光反应，分别使用波长大于320 nm或大于390 nm的光，以主要产生交联或单加合物。将受损的DNA转染到大肠杆菌后，对其失活和诱变情况进行评分。在指示培养基上出现浅蓝色或无色噬菌斑表明病毒DNA的lac插入片段发生了突变。对两种补骨脂素光处理结果的比较支持了这样一种观点，即交联比单加合物更具致死性和诱变性。大量β-半乳糖苷酶部分或完全缺陷的突变克隆通过噬菌斑纯化和扩增。通过双脱氧链终止法对每个克隆的病毒DNA进行测序。所有观察到的碱基对变化都定位到lac启动子区域，包括3个转换、14个颠换和6个单碱基对移码突变。主要的突变是T.A→G.C颠换。
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