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A General Strategy for Engineering Noncanonical Amino Acid Dependent Bacterial Growth.
- Source :
-
Journal of the American Chemical Society [J Am Chem Soc] 2019 Oct 16; Vol. 141 (41), pp. 16213-16216. Date of Electronic Publication: 2019 Oct 04. - Publication Year :
- 2019
-
Abstract
- Synthetic auxotrophy in which bacterial viability depends on the presence of a synthetic amino acid provides a robust strategy for the containment of genetically modified organisms and the development of safe, live vaccines. However, a simple, general strategy to evolve essential proteins to be dependent on synthetic amino acids is lacking. Using a temperature-sensitive selection system, we evolved an Escherichia coli ( E. coli ) sliding clamp variant with an orthogonal protein-protein interface, which contains a Leu273 to p -benzoylphenyl alanine (pBzF) mutation. The E. coli strain with this variant DNA clamp has a very low escape frequency (<10 <superscript>-10</superscript> ), and its growth is strictly dependent on the presence of pBzF. This selection strategy can be generally applied to create ncAA dependence of other organisms with DNA clamp homologues.
Details
- Language :
- English
- ISSN :
- 1520-5126
- Volume :
- 141
- Issue :
- 41
- Database :
- MEDLINE
- Journal :
- Journal of the American Chemical Society
- Publication Type :
- Academic Journal
- Accession number :
- 31580059
- Full Text :
- https://doi.org/10.1021/jacs.9b08491