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在聚合酶链式反应(PCR)过程中同时使用MutS和RecA抑制非特异性扩增

Simultaneous Use of MutS and RecA for Suppression of Nonspecific Amplification during PCR.

作者信息

Fukui Kenji, Kuramitsu Seiki

机构信息

RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan ; Science and Technology Entrepreneurship Laboratory, Osaka University, Suita, Osaka 565-0871, Japan ; Department of Biological Sciences, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan.

出版信息

J Nucleic Acids. 2013;2013:823730. doi: 10.1155/2013/823730. Epub 2013 Jul 21.

DOI:10.1155/2013/823730
PMID:23970956
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3736406/
Abstract

Thermus thermophilus MutS, a thermostable mismatch-recognizing protein, is utilized in PCR to suppress nonspecific amplification by preventing synthesis from mismatched primers. T. thermophilus RecA also decreases nonspecific amplification by promoting proper hybridization between the primer and template. We observed that MutS and RecA function under the same reaction conditions and that MutS and RecA do not preclude each other. Furthermore, there were some DNA sequences for which only one of the 2 proteins effectively suppressed nonspecific amplification. The simultaneous use of MutS and RecA is a more attractive error-suppressing technique than the use of either of the 2 proteins alone.

摘要

嗜热栖热菌MutS是一种热稳定的错配识别蛋白,在聚合酶链式反应(PCR)中用于通过阻止错配引物的合成来抑制非特异性扩增。嗜热栖热菌RecA也通过促进引物与模板之间的正确杂交来减少非特异性扩增。我们观察到MutS和RecA在相同反应条件下起作用,并且MutS和RecA并不相互排斥。此外,存在一些DNA序列,对于这些序列,两种蛋白质中只有一种能有效抑制非特异性扩增。与单独使用这两种蛋白质中的任何一种相比,同时使用MutS和RecA是一种更具吸引力的错误抑制技术。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df10/3736406/b9c8bec64b6c/JNA2013-823730.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df10/3736406/e2e51903d381/JNA2013-823730.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df10/3736406/b9c8bec64b6c/JNA2013-823730.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df10/3736406/e2e51903d381/JNA2013-823730.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df10/3736406/b9c8bec64b6c/JNA2013-823730.002.jpg

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本文引用的文献

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Thermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR).热稳定错配识别蛋白 MutS 在聚合酶链反应 (PCR) 中抑制非特异性扩增。
Int J Mol Sci. 2013 Mar 21;14(3):6436-53. doi: 10.3390/ijms14036436.
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NIH Image to ImageJ: 25 years of image analysis.NIH 图像到 ImageJ:25 年的图像分析。
Nat Methods. 2012 Jul;9(7):671-5. doi: 10.1038/nmeth.2089.
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Multiplex PCR: use of heat-stable Thermus thermophilus RecA protein to minimize non-specific PCR products.多重聚合酶链反应:使用耐热嗜热栖热菌RecA蛋白以减少非特异性聚合酶链反应产物。
Nucleic Acids Res. 2005 Aug 8;33(14):e126. doi: 10.1093/nar/gni111.
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Homologous genetic recombination as an intrinsic dynamic property of a DNA structure induced by RecA/Rad51-family proteins: a possible advantage of DNA over RNA as genomic material.同源基因重组作为由RecA/Rad51家族蛋白诱导的DNA结构的一种内在动态特性:DNA作为基因组物质相对于RNA的一种可能优势。
Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8425-32. doi: 10.1073/pnas.111005198.
9
The crystal structure of DNA mismatch repair protein MutS binding to a G x T mismatch.DNA错配修复蛋白MutS与G×T错配结合的晶体结构。
Nature. 2000 Oct 12;407(6805):711-7. doi: 10.1038/35037523.
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Crystal structures of mismatch repair protein MutS and its complex with a substrate DNA.错配修复蛋白MutS的晶体结构及其与底物DNA的复合物
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