Alanine-stretch scanning mutagenesis: a simple and efficient method to probe protein structure and function
Open Access
- 15 January 1997
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 25 (2) , 447-448
- https://doi.org/10.1093/nar/25.2.447
Abstract
We have developed a foolproof method to substitute a stretch of residues by alanines. After the introduction of aPstI site by IPCR, thus creating two alanine codons, additional codons are introduced at this site through the use of an 'alanine-stretch cartridge'. These cartridges comprise an antibiotic resistance gene flanked on both sides by alanine codons. Excision of the resistance gene byPvuII then yields the correct insertion of codons. The method is both highly reliable and flexible and should be of general use.Keywords
This publication has 4 references indexed in Scilit:
- Site-directed mutagenesis of double-stranded DNA by the polymerase chain reactionGene, 1994
- Inactivation of Antibiotics and the Dissemination of Resistance GenesScience, 1994
- Crystal structure of Escherichia coli TEM1 β‐lactamase at 1.8 Å resolutionProteins-Structure Function and Bioinformatics, 1993
- High-Resolution Epitope Mapping of hGH-Receptor Interactions by Alanine-Scanning MutagenesisScience, 1989