Isolation and characterization of Df(2R)BSC259
Stacey Christensen and Kevin Cook
Bloomington Stock Center
Indiana University
Df(2R)BSC259 was isolated as a FLP recombinase-induced recombination event involving P{XP}toud07714 and PBac{RB}e00138. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, y1 w1118; P{XP}toud07714/PBac{RB}e00138 males crossed to w1118; P{hs-hid}2, wgSp-1/CyO females. These 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}BSC259 from the segment of P{XP}toud07714 to the left of its FRT site and the segment of PBac{RB}e00138 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(2R)BSC259 predicted from the Release 4 genomic coordinates of the transposable element insertions sites are 48A3;48C4. The presence of a deletion was confirmed cytologically, though the breakpoints were not analyzed in detail. It failed to complement wal02516.