From: kercook@XXXX Subject: Isolation and characterization of Df(3L)BSC736 Date: 28 February 2009 17:34:13 GMT To: flybase-cambridgeXXXX, sjchristXXXX, ruacookXXXX, kaufmanXXXX Isolation and characterization of Df(3L)BSC736 Stacey Christensen, Kim Cook and Kevin Cook Bloomington Stock Center Indiana University Df(3L)BSC736 was isolated as a FLP recombinase-induced recombination event involving P{XP}Gad1d10811 and PBac{WH}CG18675f03146. 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}Gad1d10811/PBac{WH}CG18675f03146 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.WH5}BSC736 from the segment of P{XP}Gad1d10811 to the left of its FRT site and the segment of PBac{WH}CG18675f03146 to the right of its FRT site. Its presence was verified using the PCR methods and primers described in Parks et al. Exelixis, Inc. determined the insertion site of PBac{WH}CG18675f03146 to be Release 3 genomic coordinate 4160334 on chromosome arm 3L. This corresponds to 64A9 on the Release 3 and Release 5 genome maps. The predicted position of P{XP}Gad1d10811 on the Release 5 map is 64A5. Consequently, the cytological breakpoints of Df(3L)BSC736 are predicted to be 64A5;64A9. Df(3L)BSC736 failed to complement RopG27, RfC40B6 and masxs76. -- 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
