Construction of Synthetic Gene Circuits in the Escherichia coli Genome

The construction of stable and functional synthetic circuits in bacteria is necessary in the areas of systems and synthetic biology. The common approach using plasmids to carry foreign genetic circuits offers convenience in genetic construction but is poor in genetic stability (e.g., variation in copy number). Genome recombination provides the stable genetic maintenance of synthetic circuits, but is often labor intensive and time consuming when the genetic circuits are complex and the DNA fragments become larger. The method introduced here is modified from that reported by Wanner’s group and is available for integration of complex genetic circuits into the Escherichia coli chromosome.