From: 	Kevin Cook <kcook@XXXX>
To: 	flybase-updates@XXXX
Cc: 	Kim Cook <ruacookXXXX>, Stacey Christensen <sjchristXXXX>
Subject: 	Isolation and characterization of Df(3R)BSC513
Date: 	Tue, 15 Apr 2008  15:24:15  -0400  ( 20:24  BST)
Isolation and characterization of Df(3R)BSC513
Stacey Christensen, Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3R)BSC513 was isolated as a FLP recombinase-induced recombination 
event involving PBac{RB}e02691 and P{XP}d08394. The deletion was 
isolated as a chromosome lacking miniwhite markers in progeny of 
w[1118]; Dr[1]/TM6C, Sb[1] females crossed to P{hsFLP}1, y[1] 
w[1118]; PBac{RB}e02691/P{XP}d08394 males. The 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}BSC513 from the segment of 
PBac{RB}e02691 to the left of its FRT site and the segment of 
P{XP}d08394 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 PBac{RB}e02691 to be Release 3 genomic coordinate 3356248 on 
chromosome arm 3R and the insertion site of P{XP}d08394 to be Release 
3 genomic coordinate 4076070 on arm 3R. The Gene Disruption project 
determined the insertion site of PBac{RB}e02691 to be Release 3 
genomic coordinate 3356415 on arm 3R and the insertion site of 
P{XP}d08394 to be Release 3 genomic coordinate 4076158 on arm 3R. The 
PBac{RB}e02691 insertion site corresponds to 84D9 on the Release 3 
and Release 5 genome maps and the P{XP}d08394 insertion site 
corresponds to 84F6 on the Release 3 and Release 5 genome maps. 
Consequently, the cytological breakpoints of Df(3R)BSC513 are 
predicted to be 84D9;84F6. Df(3R)BSC513 failed to complement grn[H10] 
and Tom34[03692].
__________________________________________________________
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  47405-7005     kcook@XXXX