Abstract
The FLP/FRT site-specific recombination system of Saccharomyces cerevisiae was expressed in stably transformed tobacco plants. The FLP protein efficiently catalyzes recombination between two directly repeated FLP recombination target (FRT) sites, deleting the sequence between them. In the constructs tested here, this deletion places the CaMV 35S promoter adjacent to a hygromycin resistance marker; transcriptional activation of the marker allows direct selection of recombination events. After crossing plants containing an integrated FLP expression construct with plants containing a FLP substrate, Fl seedlings can be selected directly for hygromycin resistance, indicating that recombination occurs at, or very early after zygote formation. Molecular analysis confirmed the expected recombination product.
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References
Andrews B, Protean G, Beatty L, Sadowski P (1985) The FLP recombinase of the 2 μ circle DNA of yeast: interaction with its target sequences. Cell 40:795–803
Bayley CC, Morgan M, Dale EC, Ow DW (1992) Exchange of gene activity in transgenic plants catalyzed by the Cre-lox site-specific recombination system. Plant Mol Biol 18:353–361
Cox MM (1983) The FLP protein of the yeast 2-μm plasmid: expression of a eukaryotic genetic recombination system in Escherichia coli. Proc Natl Acad Sci USA 80:4223–4227
Cox MM (1988) FLP site-specific recombination system of Saccharomyces cerevisiae. In: Kucherlapati R, Smith GR (eds) Genetic Recombination. American Society for Microbiology, Washington, DC, pp 429–443
Dale EC, Ow DW (1990) Intra- and intermolecular site-specific recombination in plant cells mediated by bacteriophage PI recombinase. Gene 91:79–85
Dale EC, Ow DW (1991) Gene transfer with subsequent removal of the selection gene from the host genome. Proc Natl Acad Sci USA 88:10558–10562
Fromm ME, Taylor LP, Walbot V (1986) Stable transformation of maize after gene transfer by electroporation. Nature 319:791–793
Futcher AB (1986) Copy-number amplification of the 2 micron circle plasmid of Saccharomyces cerevisiae. J Theor Biol 119:197–204
Golic KG (1991) Site-specific recombination between homologous chromosomes in Drosophila. Science 252:958–961
Golic KG, Lindquist S (1989) The FLP recombinase of yeast catalyzes site-specific recombination in the Drosophila genome. Cell 59:499–509
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes to plants. Science 227:1229–1231
Huang L-C, Wood EA, Cox MM (1991) A bacterial model system for chromosomally targeting. Nucleic Acids Res 19:443–448
Koncz C, Schell J (1986) The promoter TL-DNA gene 5 controls the tissue-specific expression of chimaeric genes carried by a novel type of Agrobacterium vector. Mol Gen Genet 204:383–396
Lloyd AM, Walbot V, Davis RW (1992) Arabidopsis and Nicotiana anthocyanin production activated by maize regulators R and CI. Science 258:1773–1775
Lloyd AM, Barnason AR, Rogers SG, Byrne MC, Fraley RT, Horsch RB (1986) Transformation of Arabidopsis thaliana with Agrobacterium tumefaciens Science 234:464–466
Marsh JL, Erfle M, Wykes EJ (1984) The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene 32:481–485
Odell J, Caimi P, Sauer B, Russell S (1990) Site-directed recombination in the genome of tobacco. Mol Gen Genet 223:369–378
O'Gorman S, Fox DT, Wahl GM (1991) Recombinase-mediated gene activation and site-specific integration in mammalian cells. Science 251:1351–1355
Russell S, Hoopes J, Odell J (1992) Directed excision of a transgene from the plant genome. Mol Gen Genet 234:49–59
Sambrook J, Fritsch FF, Maniatis T (1989) Molecular cloning: a laboratory manual. 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Schardl CL, Byrd AD, Benzion G, Altschuler MA, Hildebrand DF, Hunt AG (1987) Design and construction of a versatile system for the expression of foreign genes in plants. Gene 61:1–11
Senecoff JF, Bruckner RC, Cox MM (1985) The FLP recombinase of the yeast 2-μm plasmid: characterization of its recombination site. Proc Natl Acad Sci USA 82:7270–7274
van den Elzen PJM, Townsend J, Lee KY, Bedbrook JR (1985) A chimaeric hygromycin resistance gene as a selectable marker in plant cells. Plant Mol Biol 5:299–302
Walbot V, Warren C (1988) Regulation of Mu element copy number in maize lines with an active or inactive Mutator transposable element system. Mol Gen Genet 211:27–34
Waldron C, Murphy EB, Roberts JL, Gustafson GD, Armour SL, Malcolm SK (1985) Resistance to hygromycin B: a new marker for plant transformation studies. Plant Mol Biol 5:103–108
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Communicated by E. Meyerowitz
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Lloyd, A.M., Davis, R.W. Functional expression of the yeast FLP/FRT site-specific recombination system in Nicotiana tabacum . Molec. Gen. Genet. 242, 653–657 (1994). https://doi.org/10.1007/BF00283419
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DOI: https://doi.org/10.1007/BF00283419