Isolation and characterization of Df(3R)BSC746
Stacey Christensen, Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3R)BSC746 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}e00916 and P{XP}d03025. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w1118; P{hs-hid}3, Dr1/TM6C, Sb1 cu1 females crossed to P{hsFLP}1, y1 w1118; PBac{RB}e00916/P{XP}d03025 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}BSC746 from the segment of PBac{RB}e00916 to the left of its FRT site and the segment of P{XP}d03025 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}d03025 to be Release 3 genomic coordinate 2231048 on chromosome arm 3R. This corresponds to 83E6 on the Release 3 and Release 5 genome maps. The predicted position of PBac{RB}e00916 on the Release 5 map is 83E2. Consequently, the cytological breakpoints of Df(3R)BSC746 are predicted to be 83E2;83E6. Df(3R)BSC746 failed to complement Df(3R)BSC681 and Df(3R)WIN11.