PMID:21609695

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Citation

Donath, MJ 2nd, Dominguez, MA and Withers, ST 3rd (2011) Development of an automated platform for high-throughput P1-phage transduction of Escherichia coli. J Lab Autom 16:141-7

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.

Links

PubMed Online version:10.1016/j.jala.2010.08.005

Keywords

Automation/methods; Bacteriophage P1/genetics; Escherichia coli/genetics; Escherichia coli/virology; Gene Knockout Techniques; Genetics, Microbial/methods; Transduction, Genetic/methods

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