PT - JOURNAL ARTICLE AU - Drew S. Tack AU - Austin C. Cole AU - R. Shroff AU - B.R. Morrow AU - Andrew D. Ellington TI - Evolving Bacterial Fitness with an Expanded Genetic Code AID - 10.1101/169409 DP - 2017 Jan 01 TA - bioRxiv PG - 169409 4099 - http://biorxiv.org/content/early/2017/07/28/169409.short 4100 - http://biorxiv.org/content/early/2017/07/28/169409.full AB - Evolution has for the most part used the canonical 20 amino acids of the natural genetic code to construct proteins. While several theories regarding the evolution of the genetic code have been proposed, experimental exploration of these theories has largely been restricted to phylogenetic and computational modeling. The development of orthogonal translation systems has allowed noncanonical amino acids to be inserted at will into proteins. We have taken advantage of these advances to evolve bacteria to accommodate a 21 amino acid genetic code in which the amber codon ambiguously encodes either 3-nitro-L-tyrosine or stop. Such an ambiguous encoding strategy recapitulates numerous models for genetic code expansion, and we find that evolved lineages first accommodate the unnatural amino acid, and then begin to evolve on a neutral landscape where stop codons begin to appear within genes. The resultant lines represent transitional intermediates on the way to the fixation of a functional 21 amino acid code.