Salganik R I, Dianov G L, Kokoza E B, Ovchinnikova L P, Kurbatov V A
Mol Biol (Mosk). 1979 May-Jun;13(3):625-32.
A method of selective modification of certain regions of the genome which may become useful for inactivation of certain genes or for directed mutagenesis is proposed. For this purpose RNA products of certain genes carrying alkylating groupings randomly distributed along the polymer were used. The RNA modified to an extent of 4--5 alkylating residues per 100 nucleotides retains the ability to specific formation of DNA--RNA hybrid complexes. The alkylating molecule is N,N,N'-tri-(beta-chlorethyl), N'-(p-formylphenyl)propylene diamine-1,3. The aliphatic alkylating functions serve for attachment to RNA. The aromatic alkylating function inactivated by the formyl grouping at the para-position of the benzene ring is used for modification of DNA after hybrid formation by reduction of formyl grouping with sodium borohydride. The covalently binding of modified RNA is exhibited to occur in only the case of T7 DNA H-chain, the one complementary to the RNA derivative. L-chain does not hybridize, nor does it undergo alkylation by the RNA product thus indicating high selectivity of alkylation within the hybrid complex.
提出了一种对基因组某些区域进行选择性修饰的方法,该方法可能对某些基因的失活或定向诱变有用。为此,使用了携带沿聚合物随机分布的烷基化基团的某些基因的RNA产物。每100个核苷酸修饰到含有4至5个烷基化残基程度的RNA保留了特异性形成DNA-RNA杂交复合物的能力。烷基化分子是N,N,N'-三-(β-氯乙基),N'-(对甲酰基苯基)丙二胺-1,3。脂肪族烷基化官能团用于与RNA连接。苯环对位的甲酰基使芳香族烷基化官能团失活,在用硼氢化钠还原甲酰基后,用于杂交形成后对DNA的修饰。经修饰的RNA的共价结合仅在与RNA衍生物互补的T7 DNA H链的情况下发生。L链不杂交,也不会被RNA产物烷基化,因此表明杂交复合物内烷基化具有高选择性。