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

摘要

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.