Date: Tue, 05 Dec 2006 14:55:14 -0500 To: flybase-updates@XXXX From: Kevin Cook <kcook@XXXX> Subject: Isolation and characterization of Df(2L)BSC227 Cc: Jill Gresens <jgresensXXXX>, Stacey Christensen <sjchristXXXX>, mdealXXXX, kaufmanXXXX Isolation and characterization of Df(2L)BSC227 Stacey Christensen and Kevin Cook Bloomington Stock Center Indiana University Df(2L)BSC227 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}CG8552[e02633] and P{XP}CG7778[d00356]. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, y[1] w[1118]; PBac{RB}CG8552[e02633]/P{XP}CG7778[d00356] males crossed to w[1118]; P{hs-hid}2, wg[Sp-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}BSC227 from the segment of PBac{RB}CG8552[e02633] to the left of its FRT site and the segment of P{XP}CG7778[d00356] 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' minus primer in the Hybrid PCR protocol in the Supplementary Methods. The cytological breakpoints of Df(2L)BSC227 predicted from the Release 4 genomic coordinates of the transposable element insertions sites are 28E8;29B1. It failed to complement Btk29A[k00206] and Pvr[5363]. __________________________________________________________ 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
