Isolation and characterization of Df(3L)BSC250
Jill Gresens, Stacey Christensen and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3L)BSC250 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}CG9134e00088 and P{XP}d01193. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w1118; Dr1/TM6C, Sb1 females crossed to P{hsFLP}1, y1 w1118; PBac{RB}CG9134e00088/P{XP}d01193 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}BSC250 from the segment of PBac{RB}CG9134e00088 to the left of its FRT site and the segment of P{XP}d01193 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}d01193 to be at Release 3 genomic coordinate 1315105 on chromosome arm 3L. The Gene Disruption project determined the insertion site of P{XP}d01193 to be at Release 3 genomic coordinate 1315208 on arm 3L. This corresponds to 61F6 on the Release 3 and 4 genome maps. The predicted position of PBac{RB}CG9134e00088 on the Release 4 genome map is 61F4. Consequently, the cytological breakpoints of Df(3L)BSC250 are predicted to be 61F4;61F6. Df(3L)BSC250 failed to complement mtacp1j4A6 and Df(3L)ED207.