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[在反应效应物寡脱氧核糖核苷酸的N-(2-羟乙基)吩嗪衍生物存在下,用短寡脱氧核糖核苷酸的烷基化衍生物对DNA单链片段进行有效的选择性修饰]

[Effective selective modification of a single-stranded fragment of DNA with alkylating derivatives of short oligodeoxyribonucleotides in the presence of reaction effectors N-(2-hydroxyethyl)phenazine derivatives of oligodeoxyribonucleotides].

作者信息

Zarytova V F, Kutiavin I V, Mamaev S V, Podyminogin M A

出版信息

Bioorg Khim. 1990 Dec;16(12):1653-60.

PMID:2090116
Abstract

Tri-, tetra-, penta- and hexanucleotides bearing a reactive 4-(N-methylamino-N-2-chloroethyl)benzylamide group can effectively and selectively modify a single-stranded DNA fragment (302 nucleotides) in the presence of effectors, N-(2-hydroxyethyl)phenazinium derivatives of oligonucleotides complementary to DNA sequences adjacent to the binding site of the reagent. The reagents investigated modify not only single-stranded but also secondary-structured DNA regions. The modification extent depends on the length of oligonucleotide parts of the reagent and effector. A gap between the two stretches associated with the target DNA prevents the effector from functioning. The substitution of an octanucleotide effector by two tetranucleotide ones only slightly reduces the modification extent with a hexanucleotide reagent. A very efficient and specific modification can be achieved by using two effectors flanking the reactive oligonucleotide derivative. The approach leads to the modification extent of up to 89% with a hexanucleotide reagent.

摘要

带有反应性4-(N-甲基氨基-N-2-氯乙基)苄酰胺基团的三核苷酸、四核苷酸、五核苷酸和六核苷酸,在效应物(与试剂结合位点相邻的DNA序列互补的寡核苷酸的N-(2-羟乙基)吩嗪鎓衍生物)存在的情况下,能够有效且选择性地修饰单链DNA片段(302个核苷酸)。所研究的试剂不仅能修饰单链DNA区域,还能修饰二级结构的DNA区域。修饰程度取决于试剂和效应物中寡核苷酸部分的长度。与靶DNA相关的两段序列之间的间隙会阻止效应物发挥作用。用两个四核苷酸效应物取代一个八核苷酸效应物,对于六核苷酸试剂来说,只会略微降低修饰程度。通过使用位于反应性寡核苷酸衍生物两侧的两个效应物,可以实现非常高效且特异的修饰。使用六核苷酸试剂时,这种方法可使修饰程度高达89%。

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