Integrative vectors for gene deletion and replacement

Marian Sektas, Magdalena Gregorowicz, Magdalena Kucharska, Ewa Jodelko
Pol J Microbiol
2013; 62 (1):
ICID: 1039011
Article type: Short communication
IC™ Value: 6.00
Abstract provided by Publisher
 


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.
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