使用耐热DNA聚合酶进行酶促聚合反应时各种dATP类似物的底物特性和掺入准确性。

Substrate property and incorporation accuracy of various dATP analogs during enzymatic polymerization using thermostable DNA polymerases.

作者信息

Kuwahara Masayasu, Suto Yoshiyuki, Minezaki Satoshi, Kitagata Rina, Nagashima Jun-ichi, Sawai Hiroaki

机构信息

Department of Applied Chemistry, Faculty of Engneering, Gunma University, 1-5-1 Tenjin-chou, Kiryu, Gunma 376-8515, Japan.

出版信息

Nucleic Acids Symp Ser (Oxf). 2006(50):31-2. doi: 10.1093/nass/nrl016.

DOI:10.1093/nass/nrl016

PMID:17150802

Abstract

DNA aptamers and DNAzymes with similar function to antibodies and enzymes can be produced by in vitro selection. They would be useful as research tools for molecular biology and as indicators of specific substances for the analysis of clinical and food samples. Furthermore, development of modified DNA molecules aimed to diversify function and improve activity has recently proceeded by introducing functionalities to these DNA molecules. Such functional modified DNA molecules with an aimed activity screened from a random sequence pool of modified DNA prepared by a polymerase reaction. To enhance potential of selection library and expand diversity of modified DNA that can be synthesized enzymatically, modified analogs of 2'-deoxyadenosine triphosphate were synthesized and their substrate properties for some thermostable DNA polymerases in polymerase chain reactions (PCR) were investigated. Modified DNAs were sequenced in order to analyze incorporation accuracy of modified dATP during PCR.

摘要

通过体外筛选可产生与抗体和酶功能相似的DNA适配体和DNA酶。它们可用作分子生物学的研究工具，以及用于临床和食品样品分析的特定物质的指示剂。此外，最近通过向这些DNA分子引入功能基团来进行旨在使功能多样化和提高活性的修饰DNA分子的开发。这种具有目标活性的功能性修饰DNA分子是从通过聚合酶反应制备的修饰DNA的随机序列库中筛选出来的。为了提高选择文库的潜力并扩大可通过酶促合成的修饰DNA的多样性，合成了2'-脱氧三磷酸腺苷的修饰类似物，并研究了它们在聚合酶链反应（PCR）中对某些耐热DNA聚合酶的底物特性。对修饰的DNA进行测序，以分析PCR过程中修饰的dATP的掺入准确性。