Chapter title |
All-Codon Mutagenesis for Structure-Function Studies of Chemotaxis Signaling Proteins
|
---|---|
Chapter number | 8 |
Book title |
Bacterial Chemosensing
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7577-8_8 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7576-1, 978-1-4939-7577-8
|
Authors |
Peter Ames, John S. Parkinson |
Abstract |
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. |
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Geographical breakdown
Country | Count | As % |
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Unknown | 4 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Professor | 1 | 25% |
Researcher | 1 | 25% |
Unknown | 2 | 50% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 1 | 25% |
Unknown | 3 | 75% |