title: A metabolic prototype for eliminating tryptophan from the genetic code.
abstract: We set out to reduce the chemical constitution of a living organism to 19 amino acids. A strain was constructed for reassigning the tryptophan codon UGG to histidine and eliminating tryptophan from Escherichia coli. Histidine codons in the gene for an essential enzyme were replaced with tryptophan codons and the restoration of catalytic activity by missense suppressor His-tRNA bearing a CCA anticodon was selected. We used automated cultivation to assess the stability of this genetic construct during evolution. Histidine to tryptophan mutation at codon 30 in the transketolase gene from yeast and its cognate suppressor tRNA were stably propagated in a tktAB deletant of E. coli over 2500 generations. The ratio of histidine misincorporation at tryptophan sites in the proteome increased from 0.0007 to 0.03 over 300 days of continuous culture. This result demonstrated that the genetic code can be forced to evolve by permanent metabolic selection.
authors: CEA, DSV, IG, Genoscope, 2 rue Gaston Crémieux, 91057 Evry Cedex, France. valerie.pezo@issb.genopole.fr
authors: V
authors: V
authors: Pezo
authors: D
authors: D
authors: Louis
authors: V
authors: V
authors: Guérineau
authors: J-P
authors: JP
authors: Le Caer
authors: L
authors: L
authors: Gaillon
authors: R
authors: R
authors: Mutzel
authors: P
authors: P
authors: Marlière
mesh_terms: D003062:Codon / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005815:Genetic Code; D006639:Histidine / Q000235:genetics*; D020125:Mutation, Missense; D014176:Protein Biosynthesis; D012354:RNA, Transfer, His; D014174:Transketolase / Q000235:genetics; D014364:Tryptophan / Q000235:genetics*
chemical_list: D003062:Codon; D012354:RNA, Transfer, His; D006639:Histidine; D014364:Tryptophan; D014174:Transketolase