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[根据复合物中烷基化结果测定寡核苷酸与核酸形成复合物参数的新方法。与平衡透析法的比较]

[New method of determining parameters of oligonucleotide complex formation with nucleic acids according to the results of alkylation in complexes. Comparison with the method of equilibrium dialysis].

作者信息

Grineva N I, Karpova G G, Pichko N P

出版信息

Mol Biol (Mosk). 1978 Jan-Feb;12(1):135-46.

PMID:345104
Abstract

Binding constants of oligonucleotides with nucleic acids can be determined using data of nucleic acids alkylation with 2',3'-O-4-(N-2-chloroethyl-N-methylamino)benzylidene oligonucleotides. The latter bind to complementary sequences of nucleic acids, the binding constants of which are close to that of oligonucleotides. The extent of alkylation allows to determinate the equilibrium concentration of the binding with small corrections by an equation given, and hence to estimate the binding constants by standard modes. The binding constants of oligoadenylate and oligocytydilate derivatives with 23S RNA and rRNA have been determined at 0 degrees and 20 degrees by the method suggested and by the method of equilibrium dialysis. The binding constants are consistent of the magnitudes, as well as of the plots of the binding constants versus the oligomer and RNA concentrations and versus the oligomer length. The data obtained indicate that alkylation within the complexes of tri-hexamers with rRNA proceeds under the condition of an established equilibrium. Limits of the method suggested are evaluated.

摘要

寡核苷酸与核酸的结合常数可以通过2',3'-O-4-(N-2-氯乙基-N-甲基氨基)亚苄基寡核苷酸对核酸进行烷基化的数据来测定。后者与核酸的互补序列结合,其结合常数与寡核苷酸的接近。烷基化程度可通过给定的方程在进行小校正后确定结合的平衡浓度,从而通过标准模式估算结合常数。已通过所建议的方法和平衡透析法在0℃和20℃下测定了寡腺苷酸和寡胞嘧啶酸衍生物与23S RNA和rRNA的结合常数。结合常数在大小以及结合常数与寡聚物和RNA浓度以及与寡聚物长度的关系图方面是一致的。所获得的数据表明,三六聚体与rRNA复合物内的烷基化是在建立平衡的条件下进行的。对所建议方法的局限性进行了评估。

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