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A Cre::FLP fusion protein recombines FRT or loxP sites in transgenic maize plants†
Article first published online: 8 JUL 2008
© 2008 Pioneer Hi-Bred International, Inc. Journal compilation © 2008 Blackwell Publishing Ltd
Plant Biotechnology Journal
Volume 6, Issue 8, pages 770–781, October 2008
How to Cite
Djukanovic, V., Lenderts, B., Bidney, D. and Lyznik, L. A. (2008), A Cre::FLP fusion protein recombines FRT or loxP sites in transgenic maize plants. Plant Biotechnology Journal, 6: 770–781. doi: 10.1111/j.1467-7652.2008.00357.x
- Issue published online: 1 SEP 2008
- Article first published online: 8 JUL 2008
- Received 30 January 2008; revised 1 May 2008; accepted 10 May 2008.
- DNA recombination;
- site-specific recombination;
- transgenic plant;
- Zea mays L
The coding sequences of Cre (site-specific recombinase from bacteriophage P1) and FLP (yeast 2-µm plasmid site-specific recombinase) were fused in frame to produce a novel, dual-function, site-specific recombinase gene. Transgenic maize plants containing the Cre::FLP fusion expression vector were crossed to transgenic plants containing either the loxP or FRT excision substrate. Complete and precise excisions of chromosomal fragments flanked by the respective target sites were observed in the F1 and F2 progeny plants. The episomal DNA recombination products were frequently lost. Non-recombined FRT substrates found in the F1 plants were recovered in the F2 generation after the Cre::FLP gene segregated out. They produced the recombination products in the F3 generation when crossed back to the FLP-expressing plants. These observations may indicate that the efficiency of site-specific recombination is affected by the plant developmental stage, with site-specific recombination being more prevalent in developing embryos. The Cre::FLP fusion protein was also tested for excisions catalysed by Cre. Excisions were identified in the F1 plants and verified in the F2 plants by polymerase chain reaction and Southern blotting. Both components of the fusion protein (FLP and Cre) were functional and acted with similar efficiency. The crossing strategy proved to be suitable for the genetic engineering of maize using the FLP or Cre site-specific recombination system.