Isolation and characterization of Df(1)BSC584
Kim Cook, Stacey Christensen and Kevin Cook
Bloomington Stock Center
Indiana University
Df(1)BSC584 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}e02417 and P{XP}d06504. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of PBac{RB}e02417/P{XP}d06504; 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}BSC584 from the segment of PBac{RB}e02417 to the left of its FRT site and the segment of P{XP}d06504 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}d06504 to be Release 3 genomic coordinate 17022641 on the X chromosome. The Gene Disruption project determined the insertion site of P{XP}d06504 to be Release 3 genomic coordinate 17022648 on the X chromosome. This corresponds to 16A1 on the Release 3 and Release 5 genome maps. The predicted position of PBac{RB}e02417 on the Release 5 map is 15F4 . Consequently, the cytological breakpoints of Df(1)BSC584 are predicted to be 15F4;16A1. It failed to complement f1.