Isolation and characterization of Df(3L)BSC117
Rachel Andrade and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3L)BSC117 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}e04444 and P{XP}d02813. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, w1118; PBac{RB}e04444 /P{XP}d02813 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}BSC117 from the segment of PBac{RB}e04444 to the left of its FRT site and the segment of P{XP}d02813 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 PBac{RB}e04444 to be at Release 3 genomic coordinate 7208899 on chromosome arm 3L and the insertion site of P{XP}d02813 to be at Release 3 genomic coordinate 7294608 on arm 3L. The Gene Disruption Project determined the insertion site of PBac{RB}e04444 to be at Release 3 genomic coordinate 7209004 on arm 3L and the insertion site of P{XP}d02813 to be at Release 3 genomic coordinate 7294515 on arm 3L. The cytological breakpoints of Df(3L)BSC117 predicted from these coordinates are 65F2;65F5; however our polytene chromosome squashes showed the breakpoints 65E9-F1;65F2-5 (the 65E8,9 doublet band and 65F5 are present; the 65F1,2 doublet band is absent). Df(3L)BSC117 failed to complement Df(3L)BCS33.