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寡聚(dG)12 - 18聚集体导致寡聚(dG)12 - 18·聚(C)型引物 - 模板的不均一性。

Oligo(dG)12-18 aggregates result in non-homogeneity of oligo(dG)12-18.poly(C) type primer-template.

作者信息

Mikke R, Zmudzka B

出版信息

Nucleic Acids Res. 1977 Apr;4(4):1111-22. doi: 10.1093/nar/4.4.1111.

Abstract

Studies on the absorption spectra of equimolar solutions of oligo(dG)12-18 and poly(C), poly(Cm) or poly(Ce) showed that only 13%, 3% and 3% of base pairs, repectively, form complexes. Upon centrifugation of oligo(dG)12-18 with a molar excess of poly(C) of poly(Cm) in an analytical and a preparative centrifuge, it was found that only a part of oligodeoxynucleotide sediments with the polynucleotide, i.e. more rapidly than oligo(dG)12-18, poly(C) or poly(Cm). Products of binding of oligo(dG)12-18 with poly(C), poly(Cm) or poly (Ce) direct the synthesis of poly (dG) by AMV reverse transcriptas in accordance with the reported characteristics of these primer-templates, as well as of the enzyme. These observations suggest that the solutions of oligo(dG)12-18 with (polyC) or its analogues, commonly used as primer-templates of RNA- and DNA-directed DNA polymerases, contain a polynucleotide, to which oligo(dG)12-18 aggregates are bound through a few nucleotides chains shorter than 12-18 residues. These chains of oligo(dG)12-18 containing the 3'-OH ends are capable of of initiating the reaction with DNA POLYMERASES.

摘要

对寡聚(dG)12 - 18与聚(C)、聚(Cm)或聚(Ce)等摩尔溶液的吸收光谱研究表明,分别只有13%、3%和3%的碱基对形成复合物。在分析型和制备型离心机中,将寡聚(dG)12 - 18与摩尔过量的聚(C)或聚(Cm)进行离心处理后发现,只有一部分寡脱氧核苷酸与多核苷酸一起沉降,即沉降速度比寡聚(dG)12 - 18、聚(C)或聚(Cm)更快。寡聚(dG)12 - 18与聚(C)、聚(Cm)或聚(Ce)的结合产物,按照这些引物 - 模板以及该酶已报道的特性,通过禽成髓细胞瘤病毒(AMV)逆转录酶指导聚(dG)的合成。这些观察结果表明,常用作RNA和DNA指导的DNA聚合酶引物 - 模板的寡聚(dG)12 - 18与(聚C)或其类似物的溶液中,含有一种多核苷酸,寡聚(dG)12 - 18聚集体通过几条短于12 - 18个残基的核苷酸链与之结合。这些含有3'-OH末端的寡聚(dG)12 - 18链能够引发与DNA聚合酶的反应。

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