Date: Thu, 22 Mar 2007  14:52:29  -0400
To: flybase-updates@XXXX
From: Kevin Cook <kcook@XXXX>
Subject: Isolation and characterization of Df(2R)BSC272
Isolation and characterization of Df(2R)BSC272
Stacey Christensen and Kevin Cook
Bloomington Stock Center
Indiana University
Df(2R)BSC272 was isolated as a FLP recombinase-induced recombination 
event involving P{XP}d05649 and PBac{RB}CG4712[e00550]. The deletion 
was isolated as a chromosome lacking miniwhite markers in progeny of 
P{hsFLP}1, y[1] w[1118]; P{XP}d05649/PBac{RB}CG4712[e00550] males 
crossed to w[1118]; P{hs-hid}2, wg[Sp-1]/CyO females. These males 
were heat shocked as larvae as described in Parks et al., Nature 
Genetics 36: 288-292, 2004 (FBrf0175003). This cross and crosses in 
preceding and succeeding generations maintained the original genetic 
background of the Exelixis insertion stocks (Thibault et al., Nature 
Genetics 36: 283-287, 2004; FBrf0175002). The recombination event 
generated the genetic element P+PBac{XP5.RB3}BSC272 from the segment 
of P{XP}d05649 to the left of its FRT site and the segment of 
PBac{RB}CG4712[e00550] to the right of its FRT site. Its presence was 
verified using the PCR methods and primers described in Parks et al. 
with the substitution of the primer 5'-CCAATGCGTTTATTTCAGGTCACG-3' 
for the RB3' plus or RB3' minus primer in the Hybrid PCR protocol in 
the Supplementary Methods. Exelixis, Inc. determined the insertion 
site of P{XP}d05649 to be at Release 3 genomic coordinate 8283844 on 
chromosome arm 2R. This corresponds to 49F10 on the Release 3 and 4 
genome maps. The predicted position of PBac{RB}CG4712[e00550] on the 
Release 4 map is 49F13. Consequently, the cytological breakpoints of 
Df(2R)BSC272 are predicted to be  49F10;49F13. It failed to complement Dp[a1].