Isolation and characterization of Df(2L)BSC115
Jennifer Deal, Megan Deal and Kevin Cook
Bloomington Stock Center
Indiana University
Df(2L)BSC115 was isolated as a FLP recombinase-induced recombination event involving P{XP}d11137 and PBac{WH}f02470. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, y1 w1118; P{XP}d11137/PBac{WH}f02470 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.WH5}BSC115 from the segment of P{XP}d11137 to the left of its FRT site and the segment of PBac{WH}f02470 to the right of its FRT site. Its presence was verified using the PCR methods and primers described in Parks et al.
Exelixis, Inc. determined the insertion site of P{XP}d11137 to be at Release 3 genomic coordinate 21732458 on chromosome arm 2L and the insertion site of PBac{WH}f02470 to be at Release 3 genomic coordinate 21757506 on arm 2L.
The Gene Disruption Project determined the insertion site of P{XP}d11137 to be at Release 3 genomic coordinate 21732520 on arm 2L and the insertion site of PBac{WH}f02470 to be at Release 3 genomic coordinate 21757504 on arm 2L. The cytological breakpoints of Df(2L)BSC115 predicted from these coordinates are 40B3;40B5. Deletion homozygotes are viable, but PCR failed to amplify 588 bp and 810 bp segments lying between P{XP}d11137 and PBac{WH}f03244 from the homozygotes using the primer pairs 5'-TGCGGACTGCTGTTTTTGCT-3'/5'-CTTTGTTGGTCTGTTTGTTGTG-3' and 5'-CAACACTCGCAATCATTTGTG-3'/5'-AGTCGTAAACGCTTCCTTCTGCCC-3', respectively.