RT Journal Article
SR Electronic
T1 Recombineering in <em>Vibrio natriegens</em>
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 130088
DO 10.1101/130088
A1 Henry H. Lee
A1 Nili Ostrov
A1 Michaela A. Gold
A1 George M. Church
YR 2017
UL http://biorxiv.org/content/early/2017/04/24/130088.abstract
AB Here, we show that λ-Red homologs found in the Vibrio-associated SXT mobile element potentiate allelic exchange in V. natriegens by 10,000-fold. Specifically, we show SXT-Beta (s065), SXT-Exo (s066), and λ-Gam proteins are sufficient to enable recombination of single- and double-stranded DNA with episomal and genomic loci. We characterize and optimize episomal oligonucleotide-mediated recombineering and demonstrate recombineering at genomic loci. We further show targeted genomic deletion of the extracellular nuclease gene dns using a double-stranded DNA cassette. Continued development of this recombination technology will advance high-throughput and large-scale genetic engineering efforts to domesticate V. natriegens and to investigate its rapid growth rate.