Alanine-stretch scanning mutagenesis: a simple and efficient method to probe protein structure and function.
AUTOR(ES)
Lefèvre, F
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=146430Documentos Relacionados
- A novel method to identify nucleic acid binding sites in proteins by scanning mutagenesis: application to iron regulatory protein.
- Topographical and functional investigation of Escherichia coli penicillin-binding protein 1b by alanine stretch scanning mutagenesis.
- Orthogonal combinatorial mutagenesis: a codon-level combinatorial mutagenesis method useful for low multiplicity and amino acid-scanning protocols
- A simple and efficient method for chemical mutagenesis of DNA.
- Oligonucleotide-directed mutagenesis as a general and powerful method for studies of protein function.