Grineva N I, Karpova G G, Krutilina A I, Li L I, Baev A A
Mol Biol (Mosk). 1978 Sep-Oct;12(5):980-7.
A method of complementarily directed alkylation with following elimination of the alkylated bases and with DNA cleavage at apurinic sites was used for specific fragmentation of glucosylyl DNA of T2 and T4 phages. It was shown that denatured glucosylyl T2 and T4 DNA's are modified by alkylating derivatives of hexaadenylate and heptauridylilate. The extent of alkylation reached the maximum and then stopped. The extent of elimination and chain cleavage corresponded to that of alkylation. Treatment of DNA after alkylation with (Ap)5ARCl under condition of saturation at 20 degrees gives 572 +/- 28 fragments from T4 DNA with 200--25,000 nucleotides long and 578 +/- 33 fragments from T2 DNA. Alkylation under condition of DNA saturation with (Ap)5ARCl at 40 degrees leads to 138 +/- 15 fragments from T4 DNA and 170 +/- 16 fragments from T2 DNA. Characteristics of the fragments obtained are given.
采用一种互补定向烷基化方法,随后消除烷基化碱基并在脱嘌呤位点切割DNA,用于T2和T4噬菌体的葡萄糖基化DNA的特异性片段化。结果表明,变性的葡萄糖基化T2和T4 DNA被六腺苷酸和七尿苷酸的烷基化衍生物修饰。烷基化程度达到最大值后停止。消除和链切割的程度与烷基化程度相对应。在20℃饱和条件下,用(Ap)5ARCl对烷基化后的DNA进行处理,从T4 DNA得到572±28个片段,长度为200 - 25,000个核苷酸,从T2 DNA得到578±33个片段。在40℃DNA用(Ap)5ARCl饱和的条件下进行烷基化,从T4 DNA得到138±15个片段,从T2 DNA得到170±16个片段。给出了所获得片段的特征。