Isolation and characterization of Df(3R)BSC635
Kim Cook, Stacey Christensen and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3R)BSC635 was isolated as a FLP recombinase-induced recombination event involving P{XP}d04917 and PBac{RB}CG6687e04451. 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; P{XP}d04917/PBac{RB}CG6687e04451 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}BSC635 from the segment of P{XP}d04917 to the left of its FRT site and the segment of PBac{RB}CG6687e04451 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}d04917 to be Release 3 genomic coordinate 10557253 on chromosome arm 3R. The Gene Disruption project determined the insertion site of P{XP}d04917 to be Release 3 genomic coordinate 10557267 on arm 3R. This corresponds to 88D2 on the Release 3 and Release 5 genome maps. The predicted position of PBac{RB}CG6687e04451 on the Release 5 map is 88E3. Consequently, the cytological breakpoints of Df(3R)BSC635 are predicted to be 88D2;88E3. Df(3R)BSC635 failed to complement effmer4, Df(3R)Exel8160 and Df(3R)Exel6172.