Alanine-stretch scanning mutagenesis: a simple and efficient method to probe protein structure and function | Zendy

Florence Lefèvre | Zendy

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Alanine-stretch scanning mutagenesis: a simple and efficient method to probe protein structure and function

Author(s) -

Florence Lefèvre

Publication year - 1997

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/25.2.447

Subject(s) - biology , alanine scanning , mutagenesis , alanine , gene , genetics , site directed mutagenesis , stop codon , computational biology , mutation , amino acid , mutant

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.

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