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

Fukui Kenji, Kuramitsu Seiki

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.

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

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.

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。