III. Using Wee-Ps in P-element replacement
For some experiments it might be advantageous to have
a particular gene tagged with a Wee-P element. If the
gene of interest already has a P-element in the vicinity
(preferably in an intron that is flanked by coding sequence),
it should be possible to replace that P-element with
a Wee-P element. The technique of P-element replacement
has been described (Sepp KJ,
Auld VJ. Conversion of lacZ enhancer trap lines to GAL4
lines using targeted transposition in Drosophila melanogaster.
Genetics. 1999 Mar; 151(3):1093-101), and in the
Davis lab such P-element replacements with Wee-Ps have
been successfully made. We replace a previously existing
P-element with a w+ starter Wee-P (making sure to use
the correct phase), and then flip out the w+.
The details of how to do such P-element replacement
crosses are highly dependent on the markers in the P-element
that is to be replaced. We recommend using the crossing
schemes in the Sepp and Auld paper (Sepp
KJ, Auld VJ. Conversion of lacZ enhancer trap lines
to GAL4 lines using targeted transposition in Drosophila
melanogaster. Genetics. 1999 Mar;151(3):1093-101)
as a starting point in designing a P-element replacement.
In our hands, the rates of each of the steps in the
crosses for P-element replacement were similar to the
values reported in the Sepp and Auld paper.
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