PT - JOURNAL ARTICLE AU - Henry H. Lee AU - Nili Ostrov AU - Michaela A. Gold AU - George M. Church TI - Recombineering in <em>Vibrio natriegens</em> AID - 10.1101/130088 DP - 2017 Jan 01 TA - bioRxiv PG - 130088 4099 - http://biorxiv.org/content/early/2017/04/24/130088.short 4100 - http://biorxiv.org/content/early/2017/04/24/130088.full 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.