Isolation and characterization of Df(2R)BSC778
Stacey Christensen, Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(2R)BSC778 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}e02152 and P{XP}CG3530d07361. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, y1 w1118; PBac{RB}e02152/P{XP}CG3530d07361 males crossed to w1118; P{hs-hid}2, wgSp-1/SM6a 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}BSC778 from the segment of PBac{RB}e02152 to the left of its FRT site and the segment of P{XP}CG3530d07361 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. Exelixis, Inc. determined the insertion site of PBac{RB}e02152 to be Release 3 genomic coordinate 18357358 on chromosome arm 2R. This corresponds to Release 5 coordinate  2R:19201313 . The insertion site of P{XP}CG3530d07361 is Release 5 genomic coordinate  2R:19283437 . Consequently, the breakpoints of Df(2R)BSC778 predicted from the Release 5 genomic coordinates of the transposable element insertion sites are  2R:19201313 ;19283437 and the cytological breakpoints predicted from these coordinates are 59D6;59D10. Df(2R)BSC778 failed to complement Df(2R)BSC661.