title: Development of an automated platform for high-throughput P1-phage transduction of Escherichia coli.
abstract: Synthetic biology depends on the ability to rapidly produce strains with improved phenotypes but is limited by the ability to rapidly produce strain collections with directed mutations. Here, we present a system capable of overcoming this limitation through automated P1-phage transductions of Escherichia coli. By combining the Keio collection of single-gene deletion E. coli mutants with P1-phage, it is possible to generate an engineered host-strain collection consisting of every possible gene deletion mutant. This strategy was tested by transducing 355 genetic markers from the Keio collection into five different host strains, and it achieved a 98% success rate. This method offers an improved mechanism for rapidly engineering collections of microbes and provides one method for rapidly deploying a broader synthetic biology effort.
authors: Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors: Michael J
authors: MJ
authors: Donath
authors: Miguel A
authors: MA
authors: Dominguez
authors: Sydnor T
authors: ST
authors: Withers