Isolation and characterization of Df(3R)BSC818
Stacey Christensen, Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3R)BSC818 was isolated as a FLP recombinase-induced recombination event involving P{XP}d10097 and PBac{WH}Dysf00491. 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}d10097/PBac{WH}Dysf00491 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.WH5}BSC818 from the segment of P{XP}d10097 to the left of its FRT site and the segment of PBac{WH}Dysf00491 to the right of its FRT site. Its presence was verified using the PCR methods and primers described in Parks et al. The breakpoints of Df(3R)BSC818 predicted from the Release 5 genomic coordinates of the transposable element insertion sites are  3R:15229135 ;15328813 and the cytological breakpoints predicted from these coordinates are 92A3;92A6. To confirm the presence of the deficiency, we showed that a 659 base pair fragment spanning the PBac{WH}CG31221f05025 (FBti0042417) insertion site could not be amplified from Df(3R)BSC818/PBac{WH}CG31221f05025 flies using primers 5'-GCAGCCGCACGTTACCTGCAGGG-3' and 5'-GCGACTTCAGCATGCCCAGATCCC C-3'.