Development of an Automated Platform for High-Throughput P1-Phage Transduction of Escherichia coli

作者：Donath Michael J II; Dominguez Miguel A; Withers Sydnor T III^*

来源：JALA, 2011, 16(2): 141-147.

DOI：10.1016/j.jala.2010.08.005

• transduction
• synthetic biology
• PI phage
• Keio collection

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 PI-phage transductions of Escherichia coli. By combining the Keio collection of single-gene deletion E. coli mutants with PI-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. (JALA 2011;16:141-7)

• 出版日期2011-4