FLP-mediated intermolecular recombination in the cytoplasm of Drosophila embryos.
We show that when a heat-shock-driven gene that encodes the yeast FLP recombinase is injected into preblastoderm Drosophila embryos, it promotes intermolecular recombination between two coinjected plasmids that bear the specific recombination target sequence, FRT. Minimal, 34-bp FRT sites in the two plasmids are sufficient for their cointegration. The reaction is efficient enough to produce detectable recombinants when one of the plasmids is present in as little as 1000 molecules per embryo. This is comparable to the concentration of unique chromosomal sites, raising the possibility that integration of injected plasmid DNA into FRT-bearing fly chromosomes may also be achievable. Since integrants might be stabilized against the reverse excision reaction if the recombinase could be provided in a sharp pulse, it is encouraging that efficient plasmid cointegration is also achieved when in vitro synthesized FLP RNA rather than DNA is injected into the embryos.
Konsolaki
M
M
Department of Cellular and Developmental Biology, Harvard University, Cambridge, MA 02138.
Sanicola
M
M
Kozlova
T
T
Liu
V
V
Arcà
B
B
Savakis
C
C
Gelbart
W M
WM
Kafatos
F C
FC
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D004330:Drosophila / Q000235:genetics*; D004625:Embryo, Nonmammalian; D005656:Fungal Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics; D011995:Recombination, Genetic; D014158:Transcription, Genetic
D005656:Fungal Proteins; D012313:RNA; D004247:DNA; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase