A simple and reliable method to conduct and monitor expression cassette integration into the Escherichia coli chromosome.
We report a method for the integration of expression cassettes into the Escherichia coli chromosome using rare and dispensable sugar degradation gene loci as sites for integration. Clones carrying successfully recombined DNA fragments in the chromosome are easily screened using a solid differential medium containing the respective sugar compound. As an example for the heterologous expression of a complex natural product biosynthesis pathway, we show the stepwise chromosomal integration of the zeaxanthin biosynthesis pathway from Pantoea ananatis into E. coli.
Albermann
Christoph
C
Institute of Microbiology, Universität Stuttgart, Stuttgart, Germany. christoph.albermann@imb.uni-stuttgart.de
Trachtmann
Natalie
N
Sprenger
Georg A
GA
D002876:Chromosomes, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods; D016254:Mutagenesis, Insertional / Q000235:genetics*; D020636:Pantoea / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism*; D024341:Xanthophylls / Q000235:genetics*; D065146:Zeaxanthins
D011994:Recombinant Proteins; D024341:Xanthophylls; D065146:Zeaxanthins