Article
Precise manipulation of chromosomes in vivo enables genome-wide codon replacement.
Science (New York, N.Y.) - 15 Jul 2011
Isaacs Farren J, Carr Peter A, Wang Harris H, Lajoie Marc J, Sterling Bram, Kraal Laurens, Tolonen Andrew C, Gianoulis Tara A, Goodman Daniel B, Reppas Nikos B, Emig Christopher J, Bang Duhee, Hwang Samuel J, Jewett Michael C, Jacobson Joseph M, Church George M
Abstract excerpt
We present genome engineering technologies that are capable of fundamentally reengineering genomes from the nucleotide to the megabase scale. We used multiplex automated genome engineering (MAGE) to site-specifically replace all 314 TAG stop codons with synonymous TAA codons in parallel across 32 Escherichia coli strains. This approach allowed us to measure individual recombination frequencies, confirm viability...
Topics
- Chromosomes, Bacterial
- Codon, Terminator
- Conjugation, Genetic
- Directed Molecular Evolution
- Escherichia coli
- Genetic Engineering
- Genome, Bacterial
- Genomic Instability
- Mutagenesis, Site-Directed