FLPase-mediated excision of PBac{RB}e02061 (4.2kb 5' of Pburs) and P{XP}d02171 (7.2kb 3' of Pburs) removes the Pburs coding region.
FLPase-mediated excision of PBac{RB}e02061 (4.2kb 5' of Pburs) and P{XP}d02171 (7.2kb 3' of Pburs) removes the Pburs coding region.
Depending on the exact genotype, 30-50% of homozygous and hemizygous pbursexc mutant pupae complete metamorphosis and eclose. 100% of these adults fail to inflate their wings and their exoskeleton does not harden and shows abnormal melanization. The remainder of the pharate adults express a spectrum of morphological defects diagnostic of failures at pupal ecdysis, such as partial head eversion, abnormally short legs and wings, and the presence of a persistent space between the end of the abdomen and the overlying puparium.
pbursexc homo- and hemizygous mutants do not display a difference in the duration of pre-ecdysis compared to controls. They do however, express a longer and more variable period between pullback and head eversion. Indeed, whereas 90-100% of wild-type animals evert their heads within 90 seconds of pullback, the majority of pbursexc hemizygotes take >90 seconds to do so, with 20-40% failing to do so.
Over 50% of pbursexc mutant adults exhibit a reduced head, with 20-40% exhibiting a severely reduced or absent head, compared to wild-type. These pharate adults also show significantly shorter legs and wings.
Transheterozygous pburspupal1/pbursexc mutants show partially expanded or incomplete wings, leg extension defects and abnormal tanning.
pbursexc; Ccapexc double mutants exhibit normal viability during the larval stages, but express severe defects in pupation. Although they express pre-ecdysis behavior, this phase rarely ends with a distinct pullback of the front end of the animal from the puparium. Instead, pullback is typically weak or absent, and pre-ecdysis transitions into a weaker pre-ecdysis-like behavior, which later resembles post-ecdysis behaviors. The vast majority of animals fail to express any head eversion behaviors within 30 minutes and the morphology of the resulting pharate adults is also severely affected, with 70-90% showing no head. The presence of Ccapt.1755 and pburs+t.1886 transgenes rescues these defects.
Pbursexc is rescued by Pburs+t.1886
Pbursexc/Pburspupal1 is rescued by Pburs+t.1886
A single copy of pburs+t.1886 rescues the delay between pullback and head eversion during ecdysis, along with the reduced or absent head, and shorter legs and wings found in pbursexc mutants.
The presence of pburs+t.1886 rescues the defects found in pburspupal1/pbursexc transheterozygotes.
