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A simple and reliable method to conduct and monitor expression cassette integration into the Escherichia coli chromosome
Author(s) -
Albermann Christoph,
Trachtmann Natalie,
Sprenger Georg A.
Publication year - 2010
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200900193
Subject(s) - escherichia coli , biology , chromosome , gene , dna , genetics , heterologous expression , heterologous , computational biology , microbiology and biotechnology , recombinant dna
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.