Site-directed integration system using a combination of mutant lox sites for Corynebacterium glutamicum.
The engineering of Corynebacterium glutamicum is important for enhanced production of biochemicals. To construct an optimal C. glutamicum genome, a precise site-directed gene integration method was developed by using a pair of mutant lox sites, one a right element (RE) mutant lox site and the other a left element (LE) mutant lox site. Two DNA fragments, 5.7 and 10.2 kb-long, were successfully integrated into the genome. The recombination efficiency of this system compared to that obtained by single crossover by homologous recombination was 2 orders of magnitude higher. Moreover, the integrated DNA remained stably maintained on removal of Cre recombinase. The Cre/mutant lox system thus represents a potentially attractive tool for integration of foreign DNA in the course of the engineering of C. glutamicum traits.
Suzuki
Nobuaki
N
Microbiology Research Group, Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, Kizugawa, Kyoto, 619-0292, Japan.
Inui
Masayuki
M
Yukawa
Hideaki
H
D001665:Binding Sites / Q000235:genetics; D048230:Corynebacterium glutamicum / Q000235:genetics* / Q000254:growth & development; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D017353:Gene Deletion; D005818:Genetic Engineering; D016680:Genome, Bacterial; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D011995:Recombination, Genetic
D004269:DNA, Bacterial; C045073:Cre recombinase; D019426:Integrases