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Fig. 4. FLP remobilization. (A) FLP remobilization technique. A donor
transposon contains a transgenic insert (red) together with a marker (1)
flanked by two FRT sites. An acceptor transposon, at a desired locus,
contains a second marker (2) and one FRT site. Remobilization of the
donor transposon by FLP results in the excision of its transgene and its
potential integration into the FRT site of the acceptor transposon.
This remobilization can be followed through changes in expression of marker 1,
such as white, that occur because of changes in position effects
(from yellow in the original site to orange in the acceptor site). Different
donor transposons, each containing different transgenes, can be targeted to
the same acceptor, thereby neutralizing position effects. (B) Split
white+ marker strategy. The white+
marker is divided into two parts: 5'-white+
(5') and 3'-white+ (3'). Neither part
can produce eye pigmentation alone (indicated in gray). Recombination between
appropriately localized recombination sites, FRT in this case,
results in white+ reconstitution and its expression
(orange). (C) Integration of the split white+
marker strategy into the FLP remobilization technique. The correct
remobilization and integration of the transgene (red) are identified by
white+ reconstitution (orange). Marker 2 (yellow)
identifies donor transgenes. (D,E) DrosDel elements
P{RS5} and P{RS3}. FLP-mediated recombination at (D)
P{RS5} and (E) P{RS3} results in chromosomal remnants,
P{RS5r} and P{RS3r}, respectively. Each contains one part of
the white+ marker. Both remnants can be reconstituted
through FLP remobilization of an appropriately designed donor transposon (see
C).
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