Isolation and characterization of Df(3L)BSC795
Stacey Christensen, Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3L)BSC795 was isolated as a FLP recombinase-induced recombination event involving P{XP}d02504 and PBac{RB}CG7375e02842. 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}d02504/PBac{RB}CG7375e02842 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}BSC795 from the segment of P{XP}d02504 to the left of its FRT site and the segment of PBac{RB}CG7375e02842 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 Gene Disruption project determined the insertion site of P{XP}d02504 to be Release 3 genomic coordinate 7828548 on arm 3L. This corresponds to Release 5 coordinate  3L:7862119 . The insertion site of PBac{RB}CG7375e02842 is Release 5 genomic coordinate 8190699 on arm 3L. Consequently, the breakpoints of Df(3L)BSC795 predicted from the Release 5 genomic coordinates of the transposable element insertion sites are  3L:7862119 ;8190699 and the cytological breakpoints predicted from these coordinates are 66A17;66B12. Df(3L)BSC795 failed to complement Nmtj1C7 and pbl3.