Isolation and characterization of Df(3R)BSC502
Stacey Christensen, Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3R)BSC502 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}simae01372 and P{XP}d07254. 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}simae01372/P{XP}d07254 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}BSC502 from the segment of PBac{RB}simae01372 to the left of its FRT site and the segment of P{XP}d07254 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}d07254 to be Release 3 genomic coordinate 25949619 on chromosome arm 3R. The Gene Disruption project determined the insertion site of P{XP}d07254 to be Release 3 genomic coordinate 25949847 on arm 3R. This corresponds to 99D8 on the Release 3 and Release 5 genome maps. The predicted position of PBac{RB}simae01372 on the Release 5 map is 99D3. Consequently, the cytological breakpoints of Df(3R)BSC502 are predicted to be 99D3;99D8. Df(3R)BSC502 failed to complement Df(3R)Exel6214.