Isolation and characterization of Df(2R)BSC133
Stacey Christensen and Kevin Cook
Bloomington Stock Center
Indiana University
Df(2R)BSC133 was isolated as a FLP recombinase-induced recombination event involving P{XP}d07544 and PBac{RB}CG12744e00948.  The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, y1 w1118; P{XP}d07544/PBac{RB}CG12744e00948 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}BSC133 from the segment of P{XP}d07544 to the left of its FRT site and the segment of PBac{RB}CG12744e00948 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. Exelixis, Inc. determined the insertion site of P{XP}d07544 to be at Release 3 genomic coordinate 4897230 on chromosome arm 2R.  The Gene Disruption project determined the insertion site of P{XP}d07544 to be at Release 3 genomic coordinate 4897229 on arm 2R. The cytological breakpoints of Df(2R)BSC133 predicted from the transposable element insertion sites are 46B4;46C1.  Df(2R)BSC133 failed to complement Df(2R)B5.