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Multiplex site-directed mutagenesis strategy including high-efficiency selection of the mutant PCR products.
- Source :
-
Biotechnology letters [Biotechnol Lett] 2007 Dec; Vol. 29 (12), pp. 1921-5. Date of Electronic Publication: 2007 Aug 09. - Publication Year :
- 2007
-
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.
Details
- Language :
- English
- ISSN :
- 0141-5492
- Volume :
- 29
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Biotechnology letters
- Publication Type :
- Academic Journal
- Accession number :
- 17687622
- Full Text :
- https://doi.org/10.1007/s10529-007-9474-7