Multiplex site-directed mutagenesis strategy including high-efficiency selection of the mutant PCR products

Varga-Orvos, Zoltán and Nagy, Zsolt and Mészáros, A. and Tamás, László and Poór, Gyula (2007) Multiplex site-directed mutagenesis strategy including high-efficiency selection of the mutant PCR products. BIOTECHNOLOGY LETTERS, 29 (12). pp. 1921-1925. ISSN 0141-5492

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Abstract

Site-directed mutagenesis is of great importance for probing the structure/function relationship of proteins. Developing our previous method (Nagy et al. Anal Biochem 324:301-303, 2004), here we report a multiplex strategy for site-directed mutagenesis using PCR in one tube to introduce a single mutation into three or more genes at the same time. DNA fragments carrying the desired mutation can be distinguished from each other in a standard antibiotic selection step of the transformed bacteria. Due to this strategy the mutagenesis procedure for several genes can be accelerated.

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