Isolation and characterization of Df(3L)BSC120
Rachel Andrade and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3L)BSC120 was isolated as a FLP recombinase-induced recombination event involving PBac{WH}f04976 and P{XP}d03723. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, w1118; PBac{WH}f04976/P{XP}d03723 females crossed to w1118; P{w+mC=hs-hid}2, wgSp-1/CyO males. These females 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}BSC120 from the segment of PBac{WH}f04976 to the left of its FRT site and the segment of P{XP}d03723 to the right of its FRT site. Its presence was verified using the PCR methods and primers described in Parks et al. Exelixis, Inc. determined the insertion site of PBac{WH}f04976 to be at Release 3 genomic coordinate 11863246 on chromosome arm 3L and the insertion site of P{XP}d03723 to be at Release 3 genomic coordinate 12039353 on arm 3L. The Gene Disruption Project determined the insertion site of PBac{WH}f04976 to be at Release 3 genomic coordinate 11863241 on arm 3L and the insertion site of P{XP}d03723 to be at Release 3 genomic coordinate 12039369 on arm 3L. The cytological breakpoints of Df(3L)BSC120 predicted from these coordinates are 68E3;68F2; however, our polytene chromosome squashes showed the breakpoints 68E3-F1;68F3-5 (68E3 and the 68F5,6 doublet band are present; the 68F1,2 doublet and 68F3 are absent). It failed to complement Df(3L)Exel6115 and Df(3L)vin7.