title: Replacement of Thymidine by a Modified Base in the Escherichia coli Genome.
abstract: Prokaryotic and eukaryotic genomic DNA is comprised of the four building blocks A, G, C, and T. We have begun to explore the consequences of replacing a large fraction or all of a nucleoside in genomic DNA with a modified nucleoside. As a first step we have investigated the possibility of replacement of T by 2'-deoxy-5-(hydroxymethyl)uridine (5hmU) in the genomic DNA of Escherichia coli. Metabolic engineering with phage genes followed by random mutagenesis enabled us to achieve approximately 75% replacement of T by 5hmU in the E. coli genome and in plasmids.
authors.affiliation: The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Angad P
authors.initials: AP
authors.lastname: Mehta
authors.affiliation: The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Han
authors.initials: H
authors.lastname: Li
authors.affiliation: The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Sean A
authors.initials: SA
authors.lastname: Reed
authors.affiliation: The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Lubica
authors.initials: L
authors.lastname: Supekova
authors.affiliation: The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Tsotne
authors.initials: T
authors.lastname: Javahishvili
authors.affiliation: The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
authors.forename: Peter G
authors.initials: PG
authors.lastname: Schultz
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D016680:Genome, Bacterial; D010957:Plasmids / Q000235:genetics; D013936:Thymidine / Q000031:analogs & derivatives* / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; C018080:5-hydroxymethyl-2'-deoxyuridine; D013936:Thymidine