Isolation and characterization of Df(1)BSC871
Kim Cook and Kevin Cook
Bloomington Stock Center
Indiana University
Df(1)BSC871 was isolated as a FLP recombinase-induced recombination event involving PBac{WH}Tim9bf06127 and P{XP}d06045. The deletion was isolated as a chromosome carrying two copies of the miniwhite marker in progeny of w1118 PBac{WH}Tim9bf06127/w1118 P{XP}d06045; MKRS, P{hsFLP}86E/+ females crossed to FM7h/Dp(2;Y)G, P{hs-hid}Y 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{XP3.WH3}BSC871 from the segment of PBac{WH}Tim9bf06127 to the left of its FRT site and the segment of P{XP}d06045 to the right of its FRT site. The breakpoints of Df(1)BSC871 predicted from the Release 5 genomic coordinates of the transposable element insertion sites are X:19511634 ;19682746 and the cytological breakpoints predicted from these coordinates are 18D7;18F2. To confirm the presence of the deficiency, we showed that a 367 base pair fragment spanning the P{SUPor-P}CG12703KG08105 insertion site could not be amplified from Df(1)BSC871/P{SUPor-P}CG12703KG08105 flies using primers 5'-GTGATAACTTGTTCGATAGCTCTTCG-3' and 5'-CTTCCTGTACTACAACTGGTGGTGG-3'. Also, we showed that a 758 base pair fragment spanning the P{EPgy2}meso18EEY07842 insertion site could not be amplified from Df(1)BSC871/P{EPgy2}meso18EEY07842 flies using primers 5'-CGTCTAGAGCGTGGCGATTGTGCG-3' and 5'-GGAATCTCAGGACGGCGGACTACGG-3'.