Date: Thu, 19 Jan 2006 11:43:17 \-0500 To: flybase-updates@XXXX From: Kevin Cook <kcook@XXXX> Subject: Isolation and characterization of Df(2R)BSC135 Cc: Stacey Christensen <sjchristXXXX>, mdealXXXX, Jill Gresens <jgresensXXXX>, kaufmanXXXX Isolation and characterization of Df(2R)BSC135 Stacey Christensen and Kevin Cook Bloomington Stock Center Indiana University Df(2R)BSC135 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}CG30127e02673 and P{XP}d10336. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, y1 w1118; PBac{RB}CG30127e02673/P{XP}d10336 males crossed to w1118; P{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.RB3}BSC135 from the segment of PBac{RB}CG30127e02673 to the left of its FRT site and the segment of P{XP}d10336 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. Exelixis, Inc. determined the insertion site of P{XP}d10336 to be at Release 3 genomic coordinate 14487297 on chromosome arm 2R. The cytological breakpoints of Df(2R)BSC135 predicted from these coordinates are 56C11-D1;56D5. It failed to complement Df(2R)BSC26 and hts01103. __________________________________________________________ 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
