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Addicting diverse bacteria to a noncanonical amino acid

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Abstract

Engineered orthogonal translation systems have greatly enabled the expansion of the genetic code using noncanonical amino acids (NCAAs). However, the impact of NCAAs on organismal evolution remains unclear, in part because it is difficult to force the adoption of new genetic codes in organisms. By reengineering TEM-1 β-lactamase to be dependent on a NCAA, we maintained bacterial NCAA dependence for hundreds of generations without escape.

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Figure 1: Characterization of NCAA dependent β-lactamase variants.
Figure 2: β-lactamase variant TEM-1.B9 maintained NCAA dependence in different bacterial species during serial culture.

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Acknowledgements

This work was supported by the following grants: National Security Science and Engineering Faculty Fellowship grant FA9550-10-1-0169 to A.D.E. Welch Foundation grant F-1654. Defense Advanced Research Projects Agency N66001-14-2-4051 to A.D.E. Air Force Office of Scientific Research grant FA9550-14-1-0089 to A.D.E. Defense Advanced Research Projects Agency HR0011-15-C0095 to A.D.E.

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Authors and Affiliations

  1. Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, Texas, USA

    Drew S Tack, Jared W Ellefson, Ross Thyer, Bo Wang, Jimmy Gollihar, Matthew T Forster & Andrew D Ellington

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  1. Drew S Tack
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  2. Jared W Ellefson
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  3. Ross Thyer
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  4. Bo Wang
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  5. Jimmy Gollihar
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  6. Matthew T Forster
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  7. Andrew D Ellington
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Contributions

D.S.T. designed and performed experiments and wrote the manuscript. J.W.E. designed experiments and wrote the manuscript. R.T. designed experiments and wrote the manuscript. B.W. screened TEM-1 variants. M.T.F. screened TEM-1 variants. J.G. performed computational analysis using the Molecular Operating Environment. A.D.E. directed experimental work and wrote the manuscript.

Corresponding author

Correspondence to Andrew D Ellington.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Figures 1–14 and Supplementary Tables 1–7. (PDF 1913 kb)

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Tack, D., Ellefson, J., Thyer, R. et al. Addicting diverse bacteria to a noncanonical amino acid. Nat Chem Biol 12, 138–140 (2016). https://doi.org/10.1038/nchembio.2002

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