Simple and Rapid Site-Specific Integration of Multiple Heterologous DNAs into the Escherichia coli Chromosome
Lauren A. Riley, Irenee C. Payne, Melissa P. Tumen-Velasquez, Adam M. Guss
Abstract
More rapid genetic tools can help accelerate strain engineering, even in advanced hosts like Escherichia coli. Here, we adapt a suite of site-specific recombinases to enable simple, rapid, and highly efficient site-specific integration of heterologous DNA into the chromosome. This utility of this system was demonstrated by sequential insertion of eight DNA methyltransferases into the E. coli chromosome, allowing plasmid DNA to be protected from restriction in Clostridium clariflavum and enabling genetic transformation of this organism. This integration system should also be highly portable into non-model organisms.
Topics & Concepts
BiologyEscherichia coliHeterologousSimple (philosophy)GeneticsEscherichia coli ProteinsChromosomeComputational biologyMolecular biologyMicrobiologyGenePhilosophyEpistemologyBacterial Genetics and BiotechnologyCRISPR and Genetic EngineeringRNA and protein synthesis mechanisms