Date: Thu, 25 Aug 2005  16:08:08  \-0500
To: flybase-updates@XXXX
From: Kevin Cook <kcook@XXXX>
Subject: Isolation and characterization of Df(3L)BSC122
Isolation and characterization of Df(3L)BSC122
Rachel Andrade and Kevin Cook
Bloomington Stock Center
Indiana University
Df(3L)BSC122 was isolated as a FLP recombinase-induced recombination
event involving PBac{RB}e01500 and P{XP}d06594.
The deletion was
isolated as a chromosome lacking miniwhite markers in progeny of
P{hsFLP}1, w1118; PBac{RB}e01500/P{XP}d06594 females crossed to
w1118; P{w+mC=hs-hid}2, wgSp-1/CyO males. These females 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}BSC122 from the segment of
PBac{RB}e01500 to the left of its FRT site and the segment of
P{XP}d06594 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 PBac{RB}e01500 to be at Release 3 genomic coordinate 1737276 on
chromosome arm 3L and the insertion site of P{XP}d06594 to be at
Release 3 genomic coordinate 1774835 on arm 3L. The Gene Disruption
Project determined the insertion site of PBac{rB}e01500 to be at
Release 3 genomic coordinate 1737279 on arm 3L and the insertion site
of P{XP}d06594 to be at Release 3 genomic coordinate 1774992 on arm
3L. The cytological breakpoints of Df(3L)BSC122 predicted from these
coordinates are 62B1;62B4. It failed to complement P{PZ}dlt04276,
Df(3L)Exel6088 and Df(3L)Aprt-1.
__________________________________________________________
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