Date: Tue, 21 Sep 2004 13:13:02 \-0500 To: rd120@XXXX From: Kevin Cook <kcook@XXXX> Subject: Info from Greg Beitel The following information accompanied stocks donated to the Bloomington Stock Center by Greg Beitel, Northwestern University (8/04). I. Information for Df(3L)sinunwu7 (FBab0038192) The sequence across the Df(3L)sinunwu7 breakpoint is acggttgggcctttt/aagtcagggtcggtc. The deletion can be followed using the primers nwu7 left (5'-CCATTCAGACAGCCAGTGACTT-3') and nwu7 right (5'-AAAAACGTGCACTACCAGAAACTG-3'). The product should be 759 bp for the nwu7 deletion chromosome and 1.4 kb for WT. II. TM6B, P{ry+t7.2=HZ2.7}3, Sb1 Tb1 ca1 Reference: Paul SM, Ternet M, Salvaterra PM, Beitel GJ.. The Na+/K+ ATPase is required for septate junction function and epithelial tube-size control in the Drosophila tracheal system. Development. 2003 Oct;130(20):4963-74. Constructed by: Greg Beitel and Alex Hirschi This balancer variant was created by hopping P{HZ2.7} (FBtp0000299) into TM6B, AntpHu Sb1 e1 Tb1 ca1 (=TM6B-Sb1; FBba0000282) from Bloomington stock 3619 (brm2 es ca1/TM6B, Sb1 Tb1 ca1). P{HZ2.7} was obtained from Bill McGinnis. The resulting insertion is P{HZ2.7}3. The balancer short name used at Bloomington will be TM6B, P{ry+t7.2=HZ2.7}3, Sb1 Tb1 ca1. III. CyO, P{w+mC=Dfd-EYFP}2 Constructed by: Greg Beitel, ZhiGuo Liang, Stephanie Ray, Marcus Yu and Heeren Patel Here is the description provided by Greg Beitel: 'This is a '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). The w+ has a variable expressivity, but is strong enough to be used in screens looking for w- flies. 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fragment was excised using NotI/BamHI and dropped into the NotI/BamHI sites of the pYellow HI-Pelican vector (Beitel unpublished ; Genbank accession number AY730637) which is derived from the pGreen H Pelican vector of Barolo et al., Biotechniques vol. 29, 2000).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 for those labs that received it prior to it being contributed to the stock centers.' P{Dfd-EYFP}2 is the insertion in CyO. The balancer short name used at Bloomington will be CyO, P{w+mC=Dfd-EYFP}2. __________________________________________________________ Kevin Cook, Ph.D. Bloomington Drosophila Stock Center Department of Biology http://flystocks.bio.indiana.edu Jordan Hall 142 Indiana University 812-856-1213 1001 E. Third St. 812-855-2577 (fax) Bloomington, IN 47405-3700 kcook@XXXX
