Estimation of Binding Constants of Peptide Nucleic Acid and Secondary-Structured DNA by Affinity Capillary Electrophoresis

被引：19

作者：

Kundu, Lal Mohan ^{[1
]}

Tsukada, Harumi ^{[1
,2
]}

Matsuoka, Yukiharu ^{[1
,2
]}

Kanayama, Naoki ^{[1
]}

Takarada, Tohru ^{[1
]}

Maeda, Mizuo ^{[1
,2
]}

机构：

[1] RIKEN Adv Sci Inst, Bioengn Lab, Wako, Saitama 3510198, Japan

[2] Univ Tokyo, Sch Frontier Sci, Dept Adv Mat Sci, Kashiwa, Chiba 2778561, Japan

来源：

ANALYTICAL CHEMISTRY | 2012年 / 84卷 / 12期

关键词：

SEQUENCE B-DNA; STRAND INVASION; MIXED-SEQUENCE; BLOCK-COPOLYMERS; DUPLEX INVASION; SNP ANALYSIS; PNA; HYBRIDIZATION; RECOGNITION; KINETICS;

D O I：

10.1021/ac301025m

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

An affinity capillary electrophoresis method was developed to determine a binding constant between a peptide nucleic acid (PNA) and a hairpin-structured DNA. A diblock copolymer composed of PNA and polyethylene glycol (PEG) was synthesized as a novel affinity probe. The base sequence of the probe's PNA segment was complementary to a hairpin-structured region of a 60-base single-stranded DNA (ssDNA). Upon applying a voltage, the DNA hairpin migrated slowly compared to a random sequence ssDNA in the presence of the PNA probe. This retardation was induced by strand invasion of the PNA into the DNA hairpin to form a hybridized complex, where the PEG segment received a large amount of hydrodynamic friction during electrophoresis. The binding constant between the PNA probe and the DNA hairpin was easily determined by mobility analysis. This simple method would be potentially beneficial in studying binding behaviors of various artificial nucleotides to natural DNA or RNA.

引用

页码：5204 / 5209

页数：6

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