Site-selective and hydrolytic two-strand scission of double-stranded DNA using Ce(IV)/EDTA and pseudo-complementary PNA
Site-selective and hydrolytic two-strand scission of double-stranded DNA using Ce(IV)/EDTA and pseudo-complementary PNA
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DOI：
10.1093/nar/gnh151
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发表时间：
2004-01-01
影响因子：
14.9
通讯作者：
Komiyama, M
中科院分区：
文献类型：
--
作者：
Yamamoto, Y;Uehara, A;Komiyama, M
通过将Ce（IV）/EDTA与两个伪互补肽核酸（pcPNAs）结合，双链DNA中的两条链在靶位点被位点选择性地水解。质粒DNA（4361 bp）或其线性化形式用作底物。当两个pcPNA侵入双链DNA时，只有两条链中的每一条中的指定部分不与对应DNA或pcPNA进行Watson-Crick碱基配对。在37 ℃和pH7.0下用Ce（IV）/EDTA处理该侵入复合物时，这两个单链部分在指定位点被选择性水解，导致双链DNA的位点选择性双链断裂。利用连接酶成功地将水解产物与外源双链DNA连接起来。这些人工系统操纵巨大DNA的潜力已经被指出。
By combining Ce(IV)/EDTA with two pseudo-complementary peptide nucleic acids (pcPNAs), both strands in double-stranded DNA were site-selectively hydrolyzed at the target site. Either plasmid DNA (4361 bp) or its linearized form was used as the substrate. When two pcPNAs invaded into the double-stranded DNA, only the designated portion in each of the two strands was free from Watson-Crick base pairing with the counterpart DNA or the pcPNA. Upon the treatment of this invasion complex with Ce(IV)/EDTA at 37degreesC and pH 7.0, both of these single-stranded portions were selectively hydrolyzed at the designated site, resulting in the site-selective two-strand scission of the double-stranded DNA. Furthermore, the hydrolytic scission products were successfully connected with foreign double-stranded DNA by using ligase. The potential of these artificial systems for manipulation of huge DNA has been indicated.