Chemical modification of Ce(IV)/EDTA-based artificial restriction DNA cutter for versatile manipulation of double-stranded DNA

被引：28

作者：

Yamamoto, Yoji ^{[1
]}

Mori, Masao ^{[1
]}

Aiba, Yuichiro ^{[1
]}

Tomita, Takafumi ^{[1
]}

Chen, Wen ^{[1
]}

Zhou, Jing-Min ^{[1
]}

Uehara, Akihiko ^{[1
]}

Ren, Yi ^{[1
]}

Kitamura, Yoshihito ^{[1
]}

Komiyama, Makoto ^{[1
]}

机构：

[1] Univ Tokyo, Res Ctr Adv Sci & Technol, Meguro Ku, Tokyo 1538904, Japan

来源：

NUCLEIC ACIDS RESEARCH | 2007年 / 35卷 / 07期

关键词：

D O I：

10.1093/nar/gkm052

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A monophosphate group was attached to the terminus of pseudo-complementary peptide nucleic acid (pcPNA), and two of thus modified pcPNAs were combined with Ce(IV)/EDTA for site-selective hydrolysis of double-stranded DNA. The site-selective DNA scission was notably accelerated by this chemical modification of pcPNAs. These second-generation artificial restriction DNA cutters (ARCUTs) differentiated the target sequence so strictly that no scission occurred even when only one DNA base-pair was altered to another. By using two of the activated ARCUTs simultaneously, DNA substrate was selectively cut at two predetermined sites, and the desired fragment was clipped and cloned. The DNA scission by ARCUT was also successful even when the target site was methylated by methyltransferase and protected from the corresponding restriction enzyme. Furthermore, potentiality of ARCUT for manipulation of huge DNA has been substantiated by site-selective scission of genomic DNA of Escherichia coli (composed of 4,600,000 bp) at the target site. All these results indicate promising applications of ARCUTs for versatile purposes.

引用

页数：8

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