FLP-Mediated Recombination of FRT Sites in the Maize Genome

• Published in: Nucleic acids research Vol. 24; no. 19; pp. 3784 - 3789
• Main Authors: Lyznik, L. Alexander, Rao, K. V., Hodges, Thomas K.
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.10.1996
• Subjects: Base Sequence; Cloning, Molecular; DNA Nucleotidyltransferases - genetics; DNA Nucleotidyltransferases - metabolism; DNA, Plant; Fungal Proteins - metabolism; Genetic Markers; Genetic Vectors; Genome, Plant; Glucuronidase - genetics; Molecular Sequence Data; Recombination, Genetic; Zea mays; Zea mays - genetics
• ISSN: 0305-1048; 1362-4962; 1362-4962
• DOI: 10.1093/nar/24.19.3784

Molecular evidence is provided for genomic recombinations in maize cells induced by the yeast FLP/FRT site-specific recombination system. The FLP protein recombined FRT sites previously integrated into the maize genome leading to excision of a selectable marker, the neo gene. NPTII activity was not observed after the successful recombination process; instead, the gusA gene was activated by the removal of the blocking DNA fragment. Genomic sequencing in the region of the FRT site (following the recombination reaction) indicated that a precise rearrangement of genomic DNA sequences had taken place. The functional FLP gene could be either expressed transiently or after stable integration into the maize genome. The efficiency of genomic recombinations was high enough that a selection for recombination products, or for FLP expression, was not required. The results presented here establish the FLP/FRT site-specific recombination system as an important tool for controlled modifications of maize genomic DNA.