Isolation and characterization of Df(1)BSC662
Kim Cook, Stacey Christensen and Kevin Cook
Bloomington Stock Center
Indiana University
Df(1)BSC662 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}e04303 and P{XP}d02233. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w1118 PBac{RB}e04303/ w1118 P{XP}d02233; 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}BSC662 from the segment of PBac{RB}e04303 to the left of its FRT site and the segment of P{XP}d02233 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. Exelixis, Inc. determined the insertion site of P{XP}d02233 to be Release 3 genomic coordinate 8228559 on the X chromosome. This corresponds to 7F1 on the Release 3 and Release 5 genome maps. The predicted position of PBac{RB}e04303 on the Release 5 map is 7D6. Consequently, the cytological breakpoints of Df(1)BSC662 are predicted to be 7D6;7F1. It failed to complement Cp36dec2-1 and otu7.