Monday, August 2, 2010
Pacific Concourse (Hyatt Regency San Francisco)
Modifying the genome and altering the metabolic flux of a bacterium is a technique commonly used to divert energy to desired metabolic products. However, the methods used to alter metabolic flux are often time consuming and difficult to scale up. Here we present an automated method to quickly and efficiently knockout genes in Escherichia coli using P1 phage transduction. This method was validated by transducing 355 genetic markers from the Keio collection into 5 different host strains, which resulted in a 98% success rate in 1755 attempts. Transduction was confirmed via PCR and growth curve analysis. Growth curve data will later be used to create a metabolic model to predict metabolic flux.