title: A novel selection marker for efficient DNA cloning and recombineering in E. coli.
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.
authors: Department of Genetics, University of North Carolina, Chapel Hill, NC, USA.
authors: Chuan-Wei
authors: CW
authors: Jang
authors: Terry
authors: T
authors: Magnuson
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004352:Drug Resistance, Microbial / Q000235:genetics; D050753:Enoyl-(Acyl-Carrier-Protein) Reductase (NADH) / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000235:genetics; D054889:Fatty Acid Synthase, Type II / Q000235:genetics; D023061:Gene Order; D005819:Genetic Markers; D010957:Plasmids / Q000235:genetics*; D011995:Recombination, Genetic; D014169:Transformation, Bacterial; D014260:Triclosan / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D004274:DNA, Recombinant; D029968:Escherichia coli Proteins; D005819:Genetic Markers; D014260:Triclosan; D050753:Enoyl-(Acyl-Carrier-Protein) Reductase (NADH); C493924:fabI protein, E coli; D054889:Fatty Acid Synthase, Type II