From: kercook@XXXX Subject: Isolation and characterization of Df(3R)BSC738 Date: 28 February 2009 17:34:29 GMT To: flybase-cambridgeXXXX, sjchristXXXX, ruacookXXXX, kaufmanXXXX Isolation and characterization of Df(3R)BSC738 Stacey Christensen, Kim Cook and Kevin Cook Bloomington Stock Center Indiana University Df(3R)BSC738 was isolated as a FLP recombinase-induced recombination event involving P{XP}TfIIFalphad01485 and PBac{RB}Edg84Ae02715. 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}TfIIFalphad01485/PBac{RB}Edg84Ae02715 males. The males were heat shocked as larvae as described in Parks et al., Nature Genetics 36: 288-292, 2004 (FBrf0175003). The recombination event generated the genetic element P+PBac{XP5.RB3}BSC738 from the segment of P{XP}TfIIFalphad01485 to the left of its FRT site and the segment of PBac{RB}Edg84Ae02715 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(3R)BSC738 predicted from the Release 5 genomic coordinates of the transposable element insertion sites are 83E5;84A1. Df(3R)BSC738 failed to complement gpp03342, Taf11 and lab14. -- Kevin Cook, Ph.D. Bloomington Drosophila Stock Center Department of Biology Indiana University 1001 E. Third St. Bloomington, IN 47405-7005 kercook@XXXX 812-856-1213 (office), 812-855-2577 (fax) http://flystocks.bio.indiana.edu
