Date: Wed, 08 Mar 2006 13:13:02 \-0500 To: flybase-updates@XXXX From: Kevin Cook <kcook@XXXX> Subject: Isolation and characterization of Df(2L)BSC144 Cc: Stacey Christensen <sjchristXXXX>, mdealXXXX, Jill Gresens <jgresens@XXXX> Isolation and characterization of Df(2L)BSC144 Stacey Christensen and Kevin Cook Bloomington Stock Center Indiana University Df(2L)BSC144 was isolated as a FLP recombinase-induced recombination event involving PBac{RB}e01839 and P{XP}d08708. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, y1 w1118; PBac{RB}e01839/P{XP}d08708 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}BSC144 from the segment of PBac{RB}e01839 to the left of its FRT site and the segment of P{XP}d08708 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}d08708 to be at Release 3 genomic coordinate 10450004 on chromosome arm 2L. The cytological breakpoints of Df(2L)BSC144 predicted from the transposable element insertions sites using release 3 coordinates are 31B1;31E5. It failed to complement chico1, trk1 and bsk1. __________________________________________________________ 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
