Reprograming the Replisome of a Semisynthetic Organism for the Expansion of the Genetic Alphabet.

ABSTRACT: Semisynthetic organisms (SSOs) created from Escherichia coli can replicate a plasmid containing an unnatural base pair (UBP) formed between the synthetic nucleosides dNaM and dTPT3 (dNaM-dTPT3) when the corresponding unnatural triphosphates are imported via expression of a nucleoside triphosphate transporter. The UBP can also be transcribed and used to translate proteins containing unnatural amino acids. However, UBPs are not well retained in all sequences, limiting the information that can be encoded, and are invariably lost upon extended growth. Here we explore the contributions of the E. coli DNA replication and repair machinery to the propagation of DNA containing dNaM-dTPT3 and show that replication by DNA polymerase III, supplemented with the activity of polymerase II and methyl-dire

SUBMITTER: Ledbetter MP 

PROVIDER: S-EPMC5793209 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature