A Novel Selection Marker for Efficient DNA Cloning and Recombineering in E. coli
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Authors: | Chuan-Wei Jang Terry Magnuson |
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Affiliation: | 1. Department of Genetics, University of North Carolina, Chapel Hill, North Carolina, United States of America.; 2. Carolina Center for Genome Sciences, University of North Carolina, Chapel Hill, North Carolina, United States of America.; 3. Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina, United States of America.; University of Massachusetts Medical School, United States of America, |
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Abstract: | Production of recombinant DNA in bacterial cells is an essential technique in molecular biology. Plasmids are usually maintained in an E. coli host by antibiotic selection. However, there are only a few antibiotic-resistance markers available in common use. Here we report the adoption of a novel selection marker, mfabI (mutant fabI) for plasmid propagation in E. coli. mfabI expands the limited repertoire of selection markers and allows for more efficient molecular manipulation and plasmid propagation in E. coli. We show that mfabI is not only an efficient plasmid selection marker, but it also possesses unique activity that may facilitate molecular manipulation of unstable sequences. Furthermore, we have incorporated mfabI in the recombineering tool kit for generating mouse gene targeting vectors and demonstrate the advantage of using mfabI-containing recombineering vectors. |
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