Single-molecular AFM probing of specific DNA sequencing using RecA-promoted homologous pairing and strand exchange

被引：37

作者：

Seong, GH ^{[1
]}

Niimi, T ^{[1
]}

Yanagida, Y ^{[1
]}

Kobatake, E ^{[1
]}

Aizawa, M ^{[1
]}

机构：

[1] Tokyo Inst Technol, Grad Sch Biosci & Biotechnol, Dept Biol Informat, Midori Ku, Yokohama, Kanagawa 2268501, Japan

来源：

ANALYTICAL CHEMISTRY | 2000年 / 72卷 / 06期

关键词：

D O I：

10.1021/ac990893h

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

The specific sequence in a linearlized double-stranded DNA target has been identified at a single-molecular level by atomic force microscopy (AFM). This was accomplished using RecA-coated, single-stranded DNA probes which were paired with a specific complementary DNA sequence in a linear double-stranded DNA target by strand-exchange reaction at a homologous sequence site with target DNA. The sites of interaction between the nucleoprotein filaments and the double-stranded DNA targets were directly visualized by AFM in solution containing 4 mM magnesium acetate. Measurements of the position of RecA-coated probes paired to individual target DNA showed that DNA probes specifically paired at their corresponding homologous target sequences. Strand exchange promoted by RecA and the visualization by AFM provided a rapid and efficient way to identify homologous sequence on a single-molecule target DNA.

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页码：1288 / 1293

页数：6

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