Date: Mon, 29 Oct 2007 12:07:21 -0400 To: flybase-updatesXXXX, Stacey Christensen <sjchristXXXX> From: Kevin Cook <kcook@XXXX> Subject: Isolation and characterization of Df(3L)BSC395 Isolation and characterization of Df(3L)BSC395 Stacey Christensen, Kimberley Cook and Kevin Cook Bloomington Stock Center Indiana University Df(3L)BSC395 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}CG6928[e01931] and P{XP}d03723. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w[1118]; Dr[1]/TM6C, Sb[1] females crossed to P{hsFLP}1, y[1] w[1118]; PBac{RB}CG6928[e01931]/P{XP}d03723 males. 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}BSC395 from the segment of PBac{RB}CG6928[e01931] to the left of its FRT site and the segment of P{XP}d03723 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}d03723 to be at Release 3 genomic coordinate 12039353 on chromosome arm 3L. The Gene Disruption project determined the insertion site of P{XP}d03723 to be at Release 3 genomic coordinate 12039369 on arm 3L. This corresponds to 68F2 on both the Release 3 and 5 genome maps. The predicted position of PBac{RB}CG6928[e01931] on the Release 5 map is 68F1. Consequently, the cytological breakpoints of Df(3L)BSC395 are predicted to be 68F1;68F2. Df(3L)BSC395 failed to complement rols[08232]. Df(3L)FDD-0170557 is a synonym for Df(3L)BSC395. __________________________________________________________ Kevin Cook, Ph.D. Bloomington Drosophila Stock Center Department of Biology http://flystocks.bio.indiana.edu Jordan Hall 142 Indiana University 812-856-1213 1001 E. Third St. 812-855-2577 (fax) Bloomington, IN 47405-7005 kcook@XXXX
