Isolation and characterization of Df(3L)BSC121
Rachel Andrade and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3L)BSC121 was isolated as a FLP recombinase-induced recombination event involving P{XP}d10431 and PBac{RB}e00237. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, w1118; P{XP}d10431/PBac{RB}e00237 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.RB3}BSC121 from the segment of P{XP}d10431 to the left of its FRT site and the segment of PBac{RB}e00237 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}d10431 to be at Release 3 genomic coordinate 160641 on chromosome arm 3L and the insertion site of PBac{RB}e00237to be at Release 3 genomic coordinate 184051 on arm 3L. The Gene Disruption Project determined the insertion site of P{XP}d10431 to be at Release 3 genomic coordinate 160825 on arm 3L and the insertion site of PBac{RB}e00237 to be at Release 3 genomic coordinate 184299 on arm 3L. The cytological breakpoints of Df(3L)BSC121 predicted from these coordinates are 61B2;61B2. It failed to complement Df(3L)Exel6084 and Df(3L)ED201, but complemented Df(3L)Exel6083 and Df(3L)ED4079.