Multiplex site-directed mutagenesis strategy including high-efficiency selection of the mutant PCR products.
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.
Laboratory of Functional Genomics, Biological Research Centre, Hungarian Academy of Sciences, P.O. Box 521, Szeged, 6701, Hungary.
Zoltán
Z
Varga-Orvos
Zsolt B
ZB
Nagy
Attila
A
Mészáros
Szabolcs
S
Kökény
Péter
P
Gergely
László
L
Tamás
Gyula
G
Poór
D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D005822:Genetic Vectors; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012641:Selection, Genetic
D004247:DNA; D004262:DNA Restriction Enzymes