Int. J. Mol. Sci. 2013, 14(3), 6436-6453; doi:10.3390/ijms14036436
Article

Thermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR)

1email, 1email, 2email, 3email and 1,2,* email
Received: 31 January 2013; in revised form: 28 February 2013 / Accepted: 11 March 2013 / Published: 21 March 2013
(This article belongs to the Special Issue Molecular Cut and Paste)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: Polymerase chain reaction (PCR)-related technologies are hampered mainly by two types of error: nonspecific amplification and DNA polymerase-generated mutations. Here, we report that both errors can be suppressed by the addition of a DNA mismatch-recognizing protein, MutS, from a thermophilic bacterium. Although it had been expected that MutS has a potential to suppress polymerase-generated mutations, we unexpectedly found that it also reduced nonspecific amplification. On the basis of this finding, we propose that MutS binds a mismatched primer-template complex, thereby preventing the approach of DNA polymerase to the 3' end of the primer. Our simple methodology improves the efficiency and accuracy of DNA amplification and should therefore benefit various PCR-based applications, ranging from basic biological research to applied medical science.
Keywords: polymerase chain reaction; DNA mismatch repair; mismatch-recognizing protein; Thermus thermophilus; Aquifex aeolicus
PDF Full-text Download PDF Full-Text [2137 KB, uploaded 19 June 2014 04:47 CEST]

Export to BibTeX |
EndNote


MDPI and ACS Style

Fukui, K.; Bessho, Y.; Shimada, A.; Yokoyama, S.; Kuramitsu, S. Thermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR). Int. J. Mol. Sci. 2013, 14, 6436-6453.

AMA Style

Fukui K, Bessho Y, Shimada A, Yokoyama S, Kuramitsu S. Thermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR). International Journal of Molecular Sciences. 2013; 14(3):6436-6453.

Chicago/Turabian Style

Fukui, Kenji; Bessho, Yoshitaka; Shimada, Atsuhiro; Yokoyama, Shigeyuki; Kuramitsu, Seiki. 2013. "Thermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR)." Int. J. Mol. Sci. 14, no. 3: 6436-6453.

Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert