Isolation and characterization of Df(1)BSC866
Kim Cook, Stacey Christensen and Kevin Cook
Bloomington Stock Center
Indiana University
Df(1)BSC866 was isolated as a FLP recombinase-induced recombination event involving P{XP}CG2256d02847 and PBac{WH}Traf6f00233. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w1118 P{XP}CG2256d02847/w1118 PBac{WH}Traf6f00233; 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{XP5.WH5}BSC866 from the segment of P{XP}CG2256d02847 to the left of its FRT site and the segment of PBac{WH}Traf6f00233 to the right of its FRT site. Its presence was verified using the PCR methods and primers described in Parks et al.. The breakpoints of Df(1)BSC866 predicted from the Release 5 genomic coordinates of the transposable element insertion sites are  X:7981026 ;8051355 and the cytological breakpoints predicted from these coordinates are 7D5;7D16. To confirm the presence of the deficiency, we showed that a 488 base pair fragment spanning the P{SUPor-P}CG2116KG00028 insertion site could not be amplified from Df(1)BSC866/P{SUPor-P}CG2116KG00028 flies using primers 5'-CGATGTATGAAAGCGGAAGGAAGCAAGC-3' and 5'-CCAATTCCCTGACCACAAATTGCG-3'. Also, a 630 base pair fragment spanning the PBac{PB}CG10959c02347 insertion site could not be amplified from Df(1)BSC866/PBac{PB}CG10959c02347 flies using primers 5'-CCGCTCACCAAGACGGTCACGGG-3' and 5'-CCGTGTGCTCCTTGCGACGATGC-3'.