Isolation and characterization of Df(3L)BSC116
Rachel Andrade and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3L)BSC116 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}e00847 and P{XP}d06681. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, w1118; PBac{RB}e00847 /P{XP}d06681 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}BSC116 from the segment of PBac{RB}e00847 to the left of its FRT site and the segment of P{XP}d06681 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}e00847 to be at Release 3 genomic coordinate 2354239 on chromosome arm 3L and the insertion site of P{XP}d06681 to be at Release 3 genomic coordinate 2534408 on arm 3L.
The Gene Disruption Project determined the insertion site of PBac{RB}e00847 to be at Release 3 genomic coordinate 2354423 on arm 3L and the insertion site of P{XP}d06681 to be at Release 3 genomic coordinate 2534411 on arm 3L. The cytological breakpoints of Df(3L)BSC116 predicted from these coordinates are 62D7;62E6; however, our polytene chromosome squashes showed the breakpoints 62E1-2;62E5-6. (The deletion forms a fusion band composed of part of the 62E1,2 doublet and all or part of 62E6.) Df(3L)BSC116 failed to complement Df(3L)Aprt-32 and Df(3L)R-G7.