Isolation and characterization of Df(3R)BSC633
Kim Cook, Stacey Christensen and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3R)BSC633 was isolated as a FLP recombinase-induced recombination event involving P{XP}Alyd01000 and PBac{RB}e03258. 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}Alyd01000/PBac{RB}e03258 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}BSC633 from the segment of P{XP}Alyd01000 to the left of its FRT site and the segment of PBac{RB}e03258 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. The cytological breakpoints of Df(3R)BSC633 predicted from the Release 5 genomic coordinates of the transposable element insertions sites are 84B2;84C3. Df(3R)BSC633 failed to complement Df(3R)ED7665. Df(3R)BSC633/TM6C, Sb1 cu1 males are sterile.