DNA unwinding assay using streptavidin-bound oligonucleotides.
DNA unwinding assay using streptavidin-bound oligonucleotides.
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DOI：
10.1186/1471-2199-7-43
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发表时间：
2006-11-28
影响因子：
--
通讯作者：
Kelman Z
中科院分区：
文献类型：
--
作者：
Shin JH;Kelman Z
解旋酶在许多细胞过程中发挥着重要作用，包括复制、转录和翻译。大多数解旋酶沿着双链的一条链转移，而取代互补链（DNA或RNA）。因此，解旋酶具有方向性。它们沿着核酸沿3‘→5’或5‘→3’方向移动。具有低活性的解旋酶或那些不能从仅包含一个单链悬垂区的底物启动双解绕的解旋酶的方向性很难确定。一种改进的测定解旋酶方向性的方法被开发出来，使用含有生物素化寡核苷酸的底物。作为概念的证明，与更传统的底物相比，底物显着提高了解旋酶活性和几种DNA解旋酶的方向性测定。此外，还开发了一种通用底物，可用于确定3‘→5’和5‘→3’解旋酶的方向性。研究表明，使用生物素-链亲和素复合物作为解旋酶底物可提高解旋酶活性和解旋酶方向性的测定。所描述的方法比目前可用的技术更简单。
Helicases play essential roles in many cellular processes including replication, transcription and translation. Most helicases translocate along one strand of the duplex while displacing the complementary strand (of either DNA or RNA). Thus, helicases have directionality. They move along nucleic acids in either the 3'→ 5' or 5'→ 3' direction. The directionality of helicases with low activity or of those that cannot initiate duplex unwinding from a substrate that contains only one single-stranded overhang region is difficult to determine. An improved assay to determine helicase directionality was developed that uses a substrate containing biotinylated oligonucleotides. As a proof of concept, it was shown that the substrates substantially improve helicase activity and directionality determination for several DNA helicases in comparison to more traditional substrates. In addition, a universal substrate that can be used to determine the directionality of both 3'→ 5' and 5'→ 3' helicases was developed. It is shown here that the use of a biotin-streptavidin complex as a helicase substrate improves helicase activity and the determination of helicase directionality. The method described is simpler that the currently available techniques.
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影响因子：
14.9
作者：
Constantinesco, F;Forterre, P;Elie, C
通讯作者：
Elie, C
影响因子：
14.9
作者：
TUTEJA, N;RAHMAN, K;FALASCHI, A
通讯作者：
FALASCHI, A
DOI：
10.1073/pnas.87.14.5363
发表时间：
1990-07-01
影响因子：
11.1
作者：
UMEZU, K;NAKAYAMA, K;NAKAYAMA, H
通讯作者：
NAKAYAMA, H
影响因子：
2.9
作者：
Morris, PD;Raney, KD
通讯作者：
Raney, KD
影响因子：
4.8
作者：
Shin, JH;Jiang, Y;Kelman, Z
通讯作者：
Kelman, Z