Isolation and characterization of Df(2L)BSC108
Jennifer Deal and Kevin Cook
Bloomington Stock Center
Indiana University
Df(2L)BSC108 was isolated as a FLP recombinase-induced recombination event involving P{XP}d04594 and PBac{RB}e02576. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, y1 w1118; P{XP}d04594/PBac{RB}e02576 males crossed to w1118; P{w+mC=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}BSC108 from the segment of P{XP}d04594 to the left of its FRT site and the segment of PBac{RB}e02576 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}d04594 to be at Release 3 genomic coordinate 6866624 on chromosome arm 2L and the insertion site of PBac{RB}e02576 to be at Release 3 genomic coordinate 6936961 on arm 2L. The Gene Disruption project determined the insertion site of P{XP}d04594 to be at Release 3 genomic coordinate 6866623 on arm 2L. The cytological breakpoints of Df(2L)BSC108 predicted from these coordinates are 27C2;27C6. It failed to complement P{SUPor-P}CG10354KG04760, P{PZ}Hrb27C02647, P{SUPor-P}CG18304KG00684 and weeES1.