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 w1118; Dr1/TM6C, Sb1 females crossed to P{hsFLP}1, y1 w1118; 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 grnH10 and Tom3403692.