From: Kevin Cook <kcook@XXXX> To: flybase-updatesXXXX, Stacey Christensen <sjchristXXXX> Subject: Isolation and characterization of Df(2L)BSC345 Date: Mon, 29 Oct 2007 12:07:18 -0400 ( 16:07 GMT) Isolation and characterization of Df(2L)BSC345 Stacey Christensen, Kimberley Cook and Kevin Cook Bloomington Stock Center Indiana University Df(2L)BSC345 was isolated as a FLP recombinase-induced recombination event involving P{XP}d03477 and PBac{RB}e02962. The deletion was isolated as a chromosome lacking miniwhite markers in progeny of P{hsFLP}1, y[1] w[1118]; P{XP}d03477/PBac{RB}e02962 males crossed to w[1118]; P{hs-hid}2, wg[Sp-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}BSC345 from the segment of P{XP}d03477 to the left of its FRT site and the segment of PBac{RB}e02962 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' plus or RB3' minus primer in the Hybrid PCR protocol in the Supplementary Methods. Exelixis, Inc. determined the insertion site of P{XP}d03477 to be at Release 3 genomic coordinate 13860811 on chromosome arm 2L. This corresponds to 34E1 on both the Release 3 and 5 genome maps. The predicted position of PBac{RB}e02962 on the Release 5 map is 34E5. Consequently, the cytological breakpoints of Df(2L)BSC345 are predicted to be 34E1;34E5. It failed to complement Df(2L)b87e25 and Df(2L)BSC252. Df(2L)FDD-0048357 is a synonym for Df(2L)BSC345. __________________________________________________________ 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
