Mapping the nicking efficiencies of nickase R.BbvCI for side-specific LNA-substituted substrates using rolling circle amplification.
Mapping the nicking efficiencies of nickase R.BbvCI for side-specific LNA-substituted substrates using rolling circle amplification.
复制标题
使用滚环扩增绘制切口酶 R.BbvCI 对侧特异性 LNA 取代底物的切口效率
DOI：
10.1038/srep32560
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发表时间：
2016-09-01
影响因子：
4.6
通讯作者：
Guan Y
中科院分区：
文献类型：
--
作者：
Wei H;Zhao G;Hu T;Tang S;Jiang J;Hu B;Guan Y
我们采用一种新的基于滚动圆扩增（RCA）的不对称解理分析方法来确定底物LNA修饰对rbbvci两个亚基解理的影响。采用两种不同的连接策略设计了两组切割圆形底物，分析了切割链和未切割链不同修饰位置对单链切割效率的影响。结果表明，LNA对修饰链和未修饰链的切割速率的影响都是位点依赖的。Nb。LNA修饰对BbvCI和Nt.BbvCI的影响不同。大部分修饰位点对这两种酶的切割均有较强的抑制作用。而底链T3位置的修饰对两种酶的活性几乎没有影响。结果表明，R.BbvCI的两个亚基之间存在密切的相互作用，而底部链的T3位置可能是一个不易被干扰的较不紧密的位置。
We used a novel asymmetric cleavage analysis method based on rolling circle amplification (RCA) to determine the effects of LNA modification of substrate on the two subunits of R.BbvCI cleavage. We designed two sets of cleavage circular substrates by using two different ligation strategies and analyzed the single strand cleavage efficiency affected by different modification positions both from the cleaved strands and the uncleaved strands. Results showed that the effects of LNA on cleavage rates of modified strands and unmodified strands were both site-dependent. The Nb.BbvCI and Nt.BbvCI were affected by LNA modification in different way. Most of the modification positions showed strong inhibition of both of these two nickases cleavage. However, the modification in T3 position of bottom strand hardly affected both of the two nickases activities. The results suggested an intimated interaction between the two subunits of R.BbvCI and the T3 position in bottom strand might be a less tight position which was hard to be disturbed.
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影响因子：
14.9
作者：
Kiesling T;Cox K;Davidson EA;Dretchen K;Grater G;Hibbard S;Lasken RS;Leshin J;Skowronski E;Danielsen M
通讯作者：
Danielsen M
影响因子：
1.6
作者：
Sasaki, Naoki;Isu, Anri;Sato, Kae
通讯作者：
Sato, Kae
影响因子：
14.9
作者：
Tolstrup, N;Nielsen, PS;Kauppinen, S
通讯作者：
Kauppinen, S
影响因子：
2.7
作者：
Chou, LS;Meadows, C;Lyon, E
通讯作者：
Lyon, E
影响因子：
15
作者：
Petersen, M;Bondensgaard, K;Jacobsen, JP
通讯作者：
Jacobsen, JP