Isolation and characterization of Df(3R)BSC528
Stacey Christensen, Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3R)BSC528 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}CG8312e01204 and P{XP}SpdSd10023. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w1118; P{hs-hid}3, Dr1/TM6C, Sb1 females crossed to P{hsFLP}1, y1 w1118; PBac{RB}CG8312e01204/P{XP}SpdSd10023 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}BSC528 from the segment of PBac{RB}CG8312e01204 to the left of its FRT site and the segment of P{XP}SpdSd10023 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. The cytological breakpoints of Df(3R)BSC528 predicted from the Release 5 genomic coordinates of the transposable element insertions sites are 85E1;85E1. Df(3R)BSC528 failed to complement Df(3R)Exel6264 and Df(3R)Exel6153.