Tat'kov S I, Tsivkovskiĭ R Iu, Baturina I I, Smirnova O Iu
Mol Biol (Mosk). 1995 Mar-Apr;29(2):301-7.
We have shown that deoxycytidine-5'-triphosphate modified by O-(4-aminobutyl)hydroxylamine in the pyrimidine ring, is effectively incorporated into DNA synthesizing in vitro, replacing deoxythymidine-5'-triphosphate or deoxycytidine-5'-triphosphate and inducing A-->G and G-->A transitions, respectively. UV spectroscopy and NMR spectroscopy have shown that the modified cytidine-5'-triphosphate is identical to N4-(4-aminobutoxy)-2'-deoxycytidine-5'-triphosphate. When the modified deoxycytidine-5'-triphosphate was inserted into DNA in vitro by DNA polymerase I of E. coli Klenow fragment, retardation sites correlating with poly-A sites (when the modified triphosphate replaced deoxythymidine-5'-triphosphate) or with poly-G sites (when it replaced deoxycytidine-5'-triphosphate) were revealed. Our data show high mutagenic effect of the modified deoxycytidine-5'-triphosphate inserted into DNA, allowing us to recommend this compound for localized static mutagenesis.
我们已经表明,在嘧啶环中经O-(4-氨基丁基)羟胺修饰的脱氧胞苷-5'-三磷酸能有效地掺入体外合成的DNA中,分别取代脱氧胸苷-5'-三磷酸或脱氧胞苷-5'-三磷酸,并分别诱导A→G和G→A转变。紫外光谱和核磁共振光谱表明,修饰的胞苷-5'-三磷酸与N4-(4-氨基丁氧基)-2'-脱氧胞苷-5'-三磷酸相同。当经修饰的脱氧胞苷-5'-三磷酸由大肠杆菌Klenow片段的DNA聚合酶I体外插入DNA时,发现了与聚A位点(当修饰的三磷酸取代脱氧胸苷-5'-三磷酸时)或聚G位点(当它取代脱氧胞苷-5'-三磷酸时)相关的延迟位点。我们的数据显示,插入DNA中的经修饰的脱氧胞苷-5'-三磷酸具有很高的诱变作用,这使我们推荐该化合物用于局部静态诱变。