Rapid and assured genetic engineering methods applied to Acinetobacter baylyi ADP1 genome streamlining

One goal of synthetic biology is to improve the efficiency and predictability of living cells by removing extraneous genes from their genomes. Here, we demonstrated improved methods for engineering the genome of the metabolically versatile and naturally transformable bacterium Acinetobacter baylyi ADP1. We used a Tn-Seq experiment in rich medium (LB) to identify genes essential for robust growth. Then, we tested the dispensability of 55 large regions of the ADP1 chromosome. We found that we could successfully delete 19 regions ranging in size from 21 to 118 kilobases that collectively account for 24.6% of the wild-type genome.