From: Kevin Cook <kcook@XXXX> To: FlyBase-Cambridge <flybase-cambridgeXXXX>, kaufmanXXXX, Stacey Christensen <sjchristXXXX>, Kim Cook <ruacookXXXX> Subject: Isolation and characterization of Df(3L)BSC632 Date: Mon, 20 Oct 2008 09:19:12 -0400 ( 14:19 BST) Isolation and characterization of Df(3L)BSC632 Stacey Christensen, Kim Cook and Kevin Cook Bloomington Stock Center Indiana University Df(3L)BSC632 was isolated as a FLP recombinase-induced recombination event involving P{XP}d06287 and PBac{RB}CG13902e01240. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of w1118; P{hs-hid}3, Dr1/TM6C, Sb1 cu1 females crossed to P{hsFLP}1, y1 w1118; P{XP}d06287/PBac{RB}CG13902e01240 males. The 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}BSC632 from the segment of P{XP}d06287 to the left of its FRT site and the segment of PBac{RB}CG13902e01240 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'-GCTTCTAAACGCTTACGCATAAACGATG-3' for the RB3' plus or RB3' minus primer in the Hybrid PCR protocol in the Supplementary Methods. The cytological breakpoints of Df(3L)BSC632 predicted from the Release 5 genomic coordinates of the transposable element insertions sites are 61C8;61D2. Df(3L)BSC632 failed to complement emc1 and Df(3L)BSC440. __________________________________________________________ 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
