Isolation and characterization of Df(1)BSC572
Stacey Christensen, Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(1)BSC572 was isolated as a FLP recombinase-induced recombination event involving P{XP}d09152 and PBac{RB}e04308. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{XP}d09152/PBac{RB}e04308; 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}BSC572 from the segment of P{XP}d09152 to the left of its FRT site and the segment of PBac{RB}e04308 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}e04308 to be Release 3 genomic coordinate 10834026 on the X chromosome. The Gene Disruption project determined the insertion site of PBac{RB}e04308 to be Release 3 genomic coordinate 10834192 on the X chromosome. This corresponds to 10A4 on the Release 3 and Release 5 genome maps. The predicted position of P{XP}d09152 on the Release 5 map is 9F8. Consequently, the cytological breakpoints of Df(1)BSC572 are predicted to be 9F8;10A4. It failed to complement v1.