title: Integrative vectors for gene deletion and replacement.
abstract: An improved method for gene deletion or replacement in Escherichia coli was developed. It employs a set of integrative vectors and two helper plasmids, as a temporary source of RecA and Flp activities. The integrative vectors combine several useful features including three different selection markers placed between two parallel oriented Flp recombinase target (FRT) sites. Each marker is flanked by two MCSs, for cloning the chosen homologous fragments of DNA to gene targeting. The vectors contain two properly oriented E. coli Chi sites for recombination enhancement. When required, selection markers can be excised from the chromosome resulting in unmarked strains.
authors.affiliation: Department of Microbiology University of Gdańisk, Gdańsk, Poland. marian.sektas@biol.ug.edu.pl
authors.forename: Marian
authors.initials: M
authors.lastname: Sektas
authors.forename: Magdalena
authors.initials: M
authors.lastname: Gregorowicz
authors.forename: Magdalena
authors.initials: M
authors.lastname: Kucharska
authors.forename: Ewa
authors.initials: E
authors.lastname: Jodelko
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D016296:Mutagenesis
chemical_list: D004269:DNA, Bacterial