title: Improved antibiotic resistance gene cassette for marker exchange mutagenesis in Ralstonia solanacearum and Burkholderia species.
abstract: Marker exchange mutagenesis is a fundamental approach to understanding gene function at a molecular level in bacteria. New plasmids carrying a kanamycin resistance gene or a trimethoprim resistance gene were constructed to provide antibiotic resistance cassettes for marker exchange mutagenesis in Ralstonia solanacearum and many antibiotic-resistant Burkholderia spp. Insertion sequences present in the flanking sequences of the antibiotic resistance cassette were removed to prevent aberrant gene replacement and polar mutation during mutagenesis in wild-type bacteria. Plasmids provided in this study would be convenient for use in gene cassettes for gene replacement in other Gram-negative bacteria.
authors.affiliation: Department of Medical Bioscience, Dong-A University, Busan 604-714, Republic of Korea.
authors.forename: Hae Young
authors.initials: HY
authors.lastname: Um
authors.forename: Eunsook
authors.initials: E
authors.lastname: Chung
authors.forename: Jai-Heon
authors.initials: JH
authors.lastname: Lee
authors.forename: Seon-Woo
authors.initials: SW
authors.lastname: Lee
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019117:Burkholderia / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D017353:Gene Deletion; D005822:Genetic Vectors; D007612:Kanamycin / Q000494:pharmacology; D016254:Mutagenesis, Insertional / Q000379:methods*; D010957:Plasmids; D043368:Ralstonia solanacearum / Q000235:genetics*; D011995:Recombination, Genetic; D012641:Selection, Genetic; D014295:Trimethoprim / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D007612:Kanamycin; D014295:Trimethoprim