All-Codon Mutagenesis for Structure-Function Studies of Chemotaxis Signaling Proteins

  • Peter Ames
  • John S. Parkinson
Part of the Methods in Molecular Biology book series (MIMB, volume 1729)


The technique of all-codon mutagenesis can generate mutants that represent all possible amino acid replacements at any particular residue in a protein. It is thus a powerful tool to probe structure-function relationships in proteins of interest. In this chapter, we describe how we used all-codon mutagenesis to obtain mutants of the Escherichia coli serine receptor Tsr with amino acid replacements at residue F373, a functionally important site in this protein. We provide general protocols for mutagenesis of a target codon in a plasmid-borne gene and for the selection and screening of the resultant mutants. These techniques should be adaptable for the study of a variety of bacterial proteins.


Chemoreceptor Chemotaxis Counter-selection Polymerase chain reaction Site-directed mutagenesis 



Our work is supported by research grant GM19559 from the National Institute of General Medical Sciences. DNA sequencing and primer synthesis were carried out by the Protein-DNA Core Facility at the University of Utah, which receives support from National Cancer Institute grant CA42014 to the Huntsman Cancer Institute.


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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Department of BiologyUniversity of UtahSalt Lake CityUSA

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