Subject: dome
Dear Rachel,
This info relates to domeless (dome). Not all of this will be published
in our Current Biology paper.
dome is the computer annotated GADFLY gene CG14226. We isolated the
dome alleles from the Gottingen X P collection. We named all alleles
using as superscript the original number used in the Gottingen X P
collection (i.e.dome367 corresponds to l(1)G0367). By plasmid rescue
we found that seven elements are inserted 400bp apart in 18E. l(1)G0321
is inserted 30bp upstream the start of LD32858 and has no zygotic nor
maternal cuticle phenotype. The other rescued P elements are inserted
in the 5' UTR at the following positions with respect to the start of
LD32858: dome405 is inserted at 20b, dome367 77 b, dome217 and
dome218 154b, dome468 268 and dome441 376b. We have other dome
alleles that have not been rescued: l(1)G0199b, l(1)G0264 and
l(1)G0282. 321, 405 and 367 are hypomorphic insertions, the others have
strong zygotic domeless phenotypes. Based on the identical phenotypes
in germ line clones of the strong alleles to germ line clones of
stat92E null alleles, they may be null.
There is a further strong allele: dome9 that was isolated from line
l(1)G0009. This is not caused by the P insertion which is in Act5C as
the phenotype can be separated from the P insertion.
We know that all the above mutants are dome alleles because the zygotic
spiracle phenotypes are rescued by UAS-dome5.1. (see below).
UAS constructs: The EST LD32858 was obtained from Research Genetics
from the embryonic 0-22 hours LD library. UAS-dome was made by an
EcoR1/Xho1 excision from pOT2a and directionally subcloned into
EcoR1/Sal1 cut pUAST. UAS-domeΔTMCYT and UAS-domeΔCYT were
similarly subcloned but using the Bgl2 and EcoRV sites within dome.
Several independent lines were tested for each experiment. One of the
UAS-dome constructs (UAS-dome5.1) rescues the spiracle phenotype even
in the absence of a GAL4 driver line, suggesting that a nearby enhancer
activates the construct in the posterior spiracles.
James