Isolation and characterization of Df(1)BSC715
Stacey Christensen, Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(1)BSC715 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}CG8924e01671 and P{XP}d04224. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w1118 PBac{RB}CG8924e01671/w1118 P{XP}d04224; MKRS, P{hsFLP}86E/+ females crossed to Binsinscy/Y males. These females 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}BSC715 from the segment of PBac{RB}CG8924e01671 to the left of its FRT site and the segment of P{XP}d04224 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'-GCTTCTAAACGCTTACGCATAAACGATG-3' for the RB3' plus or RB3' minus primer in the Hybrid PCR protocol in the Supplementary Methods. The cytological breakpoints of Df(1)BSC715 predicted from the Release 5 genomic coordinates of the transposable element insertions sites are 14A1;14B14. The presence of a deletion was confirmed cytologically, though the breakpoints were not analyzed in detail.