Subject: w- version of CyO, P{Dfd-EYFP}2
The following information was provided to the Bloomington Stock Center by
Greg Beitel, Northwestern University (10/04) to describe the isolation of a
w- derivative of CyO, P{w+mC=Dfd-EYFP}2.
'Information for CyO dfd-eYFP w-
Constructed by: Greg Beitel, Matthew Slovitt, ZhiGuo Liang, Stephanie Ray,
Marcus Yu, Heeren Patel
This is a 'w-,' 'direct drive' (i.e. not Gal4/UAS) fluorescent balancer
with strong eYFP expression in the head region visible from stage late 14
and continuing through adulthood. In late embryos and larva there is
expression in the posterior spiracles and to some extent the denticle
belts. The YFP is easily scorable by compound or dissecting
microscope. In adults the proboscis is scorable. The eYFP can be stained
immunohistochemically using a standard 25 min 4% formaldehyde fix and a
the 3E6 mouse anti-GFP monoclonal (Molecular Probes). This balancer
chromosome is 'w-' because the w+ transformation marker was inactivated by
a gamma ray induced mutation that does not seem to significantly affect
YFP expression. The nature of the mutation that inactivates the w+ is not
know. Alternative w- revertant strains are available from G. Beitel if
this isolate has collateral damage that deletes your favorite gene as well
as the w+.
The dfd-HIYFP transposon was made using the autoregulatory element of the
deformed promotor and contains the 2.7 kb genomic Xba fragment named 'Hz2.7
REV' that had been cloned by Bergson and McGinnis (EMBO vol. 9 1990) into
pBluescript. The Hz2.7REV fragment was excised using NotI/BamHI and
dropped into the NotI/BamHI sites of the pYellow HI-Pelican vector (Beitel
unpublished ; Genbank accession number AY730637) which is derived from the
pGreen H Pelican vector of Barolo et al., Biotechniques vol. 29,
2000). The pYellow HI-Pelican contains eYFP (Clontech) rather than the
original eGFP, and a small intron was added between the transcription
start site and the eYFP coding sequence. The intron was added to test if
splicing would improve expression, but this intron does not appear to
significantly improve expression compared to an intronless
construct (G. Beitel unpublished). The original isolation name of this
line was  dfd::HIYFP4_MWS5 .'